Medical NGF antagonists
阅读说明：本技术 医用ngf拮抗剂 (Medical NGF antagonists ) 是由 H·詹 L·阮 R·陈 F·钱 S·J·李 Q·楚 于 2020-03-20 设计创作，主要内容包括：提供了与来自伴生动物物种、结合NGF的TrkA ECD多肽相关的各种实施方案。此类多肽可用于治疗与诸如犬类、猫类和马类等伴生动物中的慢性疼痛和/或炎性疼痛相关的NGF诱导的病症的方法。(Various embodiments are provided relating to TrkA ECD polypeptides from companion animal species that bind NGF. Such polypeptides are useful in methods of treating NGF-induced disorders associated with chronic pain and/or inflammatory pain in companion animals such as canines, felines, and equines.)
1. A contiguous polypeptide comprising at least one extracellular domain of a TrkA polypeptide (TrkAECD polypeptide) from an accompanying animal species and a fusion partner.
2. The continuous polypeptide of claim 1, wherein the continuous polypeptide is less than 5x10 as measured by biofilm layer interferometry-6M, less than 1x10-6M, less than 5x10-7M, less than 1x10-7M, less than 5x10-8M, less than 1x10-8M, less than 5x10- 9M, less than 1x10-9M, less than 5x10-10M, less than 1x10-10M, less than 5x10-11M, less than 1x10-11M, less than 5x10-12M or less than 1x10-12The dissociation constant (Kd) of M binds to the NGF polypeptide.
3. The continuous polypeptide of claim 2, wherein the NGF polypeptide is a human NGF polypeptide, a canine NGF polypeptide, a feline NGF polypeptide, or an equine polypeptide.
4. The continuous polypeptide of any one of the preceding claims, wherein the continuous polypeptide reduces NGF signaling in the companion animal species.
5. The continuous polypeptide of any one of the preceding claims wherein the companion animal species is canine, feline, or equine.
6. The contiguous polypeptide of any one of the preceding claims, wherein the amino acid sequence of the TrkA ECD polypeptide is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence of seq id no: SEQ ID NO 2, SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 5, SEQ ID NO 7, SEQ ID NO 8, SEQ ID NO 9, SEQ ID NO 10, SEQ ID NO 12, SEQ ID NO 13, SEQ ID NO 14 or SEQ ID NO 15.
7. The continuous polypeptide of any one of the preceding claims, wherein the TrkA ECD polypeptide comprises:
a) a cysteine at a position corresponding to position 7 and position 89 of SEQ ID NO 2,3, 4, 7, 8, 9, 12, 13 or 14; or
b) A cysteine at a position corresponding to position 5 and position 87 of SEQ ID NO 5, SEQ ID NO 10 or SEQ ID NO 15.
8. The continuous polypeptide of any one of the preceding claims, wherein the TrkA ECD polypeptide comprises:
a) a cysteine at position 7 and position 89 of SEQ ID NO 2,3, 4, 7, 8, 9, 12, 13 or 14; or
b) Cysteines at positions 5 and 87 of SEQ ID NO 5, 10 or 15.
9. The contiguous polypeptide of any of the preceding claims, wherein the TrkA ECD polypeptide comprises at least one N-linked glycosylation site that is not present in a corresponding wild-type TrkA ECD polypeptide, wherein the N-linked glycosylation site comprises the sequence asparagine-xaa-serine or asparagine-xaa-threonine, wherein xaa is any amino acid other than proline, and wherein one N-linked glycosylation site does not overlap with another N-linked glycosylation site.
10. The contiguous polypeptide of any preceding claim, wherein the TrkA ECD comprises at least one N-linked glycosylation site at one or more positions selected from:
a) 3, 4, 8, 9, 13 or 14 amino acid positions 6-8, 31-33, 84-86, 85-87, 86-88, 88-90, 90-92, 92-94 and/or 94-96; or
b) Amino acid positions 4-6, 29-31, 82-84, 83-85, 84-86, 86-88, 89-90, 90-92 and/or 92-94 of SEQ ID NO 5, SEQ ID NO 10 or SEQ ID NO 15.
11. The continuous polypeptide of any one of the preceding claims, wherein the TrkA ECD polypeptide comprises:
a) an amino acid other than proline at an amino acid position corresponding to position 30 and/or position 85 of SEQ ID NO 3, 4, 8, 9, 13 or 14; and/or
b) An amino acid other than proline at an amino acid position corresponding to position 28 or position 83 of SEQ ID NO 5, SEQ ID NO 10 or SEQ ID NO 15.
12. The continuous polypeptide of any one of the preceding claims, wherein the TrkA ECD polypeptide comprises:
a) valine, glutamic acid, alanine or isoleucine at an amino acid position corresponding to position 30 and/or position 85 of SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 8, SEQ ID NO 9, SEQ ID NO 13 or SEQ ID NO 14; and/or
b) Valine, glutamic acid, alanine or isoleucine at an amino acid position corresponding to position 28 or 83 of SEQ ID NO 5, 10 or 15.
13. The continuous polypeptide of any one of the preceding claims, wherein the TrkA ECD polypeptide comprises one or more amino acid modifications listed in Table A:
wherein X indicates any amino acid other than proline (e.g. E, V, A or I);
table B:
wherein X indicates any amino acid other than proline (e.g. E, V, A or I); and/or
Table C:
wherein X indicates any amino acid other than proline (e.g. E, V, A or I).
14. The continuous polypeptide of any one of the preceding claims, wherein the TrkA ECD polypeptide comprises an amino acid sequence selected from the group consisting of: SEQ ID NO 2, SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 5, SEQ ID NO 7, SEQ ID NO 8, SEQ ID NO 9, SEQ ID NO 10, SEQ ID NO 12, SEQ ID NO 13, SEQ ID NO 14, SEQ ID NO 15, SEQ ID NO 25, SEQ ID NO 26, SEQ ID NO 27, SEQ ID NO 28, SEQ ID NO 29, SEQ ID NO 30, SEQ ID NO 31, SEQ ID NO 32 and SEQ ID NO 33.
15. The contiguous polypeptide of any of the preceding claims, wherein the contiguous polypeptide comprises a linker.
16. The continuous polypeptide of claim 15, wherein the linker comprises an amino acid sequence selected from the group consisting of: G. GG, GGG, S, SS, SSS, GS, GSGS (SEQ ID NO:143), GSGSGS (SEQ ID NO:144), GGS, GGSGGS (SEQ ID NO:145), GGSGGSGGS (SEQ ID NO:146), GGGS (SEQ ID NO:147), GGGSGGGS (SEQ ID NO:148), GGGSGGGSGGGS (SEQ ID NO:149), GSS, GSSGSS (SEQ ID NO:150), GSSGSSGSS (SEQ ID NO:151), GGSS (SEQ ID NO:152), GGSSGGSS (SEQ ID NO:153), GGSSGGSSGGS (SEQ ID NO:154), SGGSGGS (SEQ ID NO:155) and SGGGSGGGS (SEQ ID NO: 156).
17. The continuous polypeptide of any one of the preceding claims, wherein the fusion partner is selected from the group consisting of an Fc polypeptide, albumin, and an albumin binding fragment.
18. The continuous polypeptide of any one of the preceding claims, wherein the fusion partner is an Fc polypeptide comprising (a) a wild-type or variant canine IgG-A, IgG-B, IgG-C or IgG-D polypeptide; (b) a wild-type or variant cat class IgG1a, IgG1b, or IgG2 polypeptide; or (c) a wild-type or variant equine IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, or IgG7 polypeptide.
19. The continuous polypeptide of any one of the preceding claims, comprising:
formula (I): TrkA ECD 1-L1-Fc;
formula (II): Fc-L1-TrkA ECD 1;
formula (III): TrkA ECD 1-L1-Fc-L2-TrkA ECD 2;
formula (IV): TrkA ECD 1-L1-TrkA ECD 2-L2-Fc; or
Formula (V): Fc-L1-TrkA ECD 1-L2-TrkA ECD 2,
wherein TrkA ECD 1 is a first TrkA ECD polypeptide, TrkA ECD 2 is a second TrkA ECD polypeptide, L1 and L2 are optional linkers, and Fc is a wild-type or variant IgG Fc polypeptide of an accompanying animal species.
20. The contiguous polypeptide of claim 19, wherein TrkA ECD 1 and TrkA ECD 2 are the same polypeptide.
21. The contiguous polypeptide of claim 19, wherein TrkA ECD 1 and TrkA ECD 2 are different polypeptides.
22. The continuous polypeptide of any one of the preceding claims, wherein the fusion partner or Fc is a variant Fc polypeptide comprising:
a) at least one amino acid modification relative to a wild-type IgG Fc polypeptide of a companion animal species, wherein the variant IgG Fc polypeptide has increased binding affinity for protein a relative to the wild-type IgG Fc polypeptide;
b) at least one amino acid modification relative to a wild-type IgG Fc polypeptide of a companion animal species, wherein the variant IgG Fc polypeptide has reduced binding affinity to C1q relative to the wild-type IgG Fc polypeptide;
c) at least one amino acid modification relative to a wild-type IgG Fc polypeptide of a companion animal species, wherein the variant IgG Fc polypeptide has reduced binding affinity to CD16 relative to the wild-type IgG Fc polypeptide;
d) a hinge region comprising at least one amino acid modification relative to a wild-type feline or equine IgG Fc polypeptide;
e) at least one amino acid substitution relative to a wild-type cat class IgG Fc polypeptide, wherein the at least one amino acid substitution is cysteine, and wherein the variant IgG Fc polypeptide is capable of forming at least one additional interchain disulfide bond relative to the wild-type cat class IgG Fc polypeptide;
f) at least one amino acid substitution relative to a wild-type IgG Fc polypeptide derived from a companion animal species, wherein the variant Fc polypeptide is capable of binding to a neonatal Fc receptor (FcRn) with increased affinity relative to the wild-type Fc polypeptide.
23. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide as measured by biofilm layer interferometryGreater than 5x10-6M, greater than 1x10-5M, greater than 5x10- 5M, greater than 1x10-4M, greater than 5x10-4M or greater than 1x10-3The dissociation constant (Kd) of M binds to C1q and/or CD 16.
24. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide that is less than 5x10 as measured by biofilm layer interferometry-6M, less than 1x10-6M, less than 5x10- 7M, less than 1x10-7M, less than 5x10-8M, less than 1x10-8M, less than 5x10-9M, less than 1x10-9M, less than 5x10-10M, less than 1x10-10M, less than 5x10-11M, less than 1x10-11M, less than 5x10-12M or less than 1x10-12The dissociation constant (Kd) of M binds to protein A.
25. The contiguous polypeptide of any of the preceding claims, comprising a variant IgG Fc polypeptide that binds to FcRn with greater affinity than the wild-type IgG Fc polypeptide, as measured by biofilm layer interference, surface plasmon resonance, or any protein-protein interaction means at a pH in the range of about 5.0 to about 6.5, as measured at a pH of about 5.0, a pH of about 5.2, a pH of about 5.5, a pH of about 6.0, a pH of about 6.2, or a pH of about 6.5.
26. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide that is less than 5x10-6M, less than 1x10-6M, less than 5x10-7M, less than 1x10-7M, less than 5x10-8M, less than 1x10-8M, less than 5x10-9M, less than 1x10-9M, less than 5x10-10M, less than 1x10-10M, less than 5x10-11M, less than 1x10-11M, less than 5x10-12M or less than 1x10-12The dissociation constant (Kd) of M binds to FcRn, as measured by biofilm layer interferometry, surface plasmon resonance, or any protein-protein interaction means at a pH in the range of about 5.0 to about 6.5, such as at a pH of about 5.0, a pH of about 5.5, a pH of about 6.0, or a pH of about 6.5.
27. The contiguous polypeptide of any of the preceding claims, comprising a variant IgG Fc polypeptide that binds to FcRn with increased affinity relative to the wild-type Fc polypeptide, and wherein the contiguous polypeptide has increased serum half-life relative to a contiguous polypeptide comprising a wild-type Fc polypeptide.
28. The continuous polypeptide of any one of the preceding claims, wherein the wild-type IgG Fc polypeptide is:
a) a canine IgG-A Fc, IgG-B Fc, IgG-C Fc, or IgG-D Fc;
b) equine IgG1Fc, IgG2Fc, IgG3 Fc, IgG4 Fc, IgG5 Fc, IgG6Fc, or IgG7 Fc; or
c) Cat class IgG1a Fc, IgG1b Fc or IgG2 Fc.
29. The continuous polypeptide of any one of the preceding claims, wherein the wild-type IgG Fc polypeptide comprises the amino acid sequence of: SEQ ID NO 34, 35, 36, 37, 38, 39, 70, 71, 72, 73, 74, 75, 76, 77, 86, 87, 88, 89 or 90.
30. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising:
a) at least one amino acid substitution relative to a wild-type cat class IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 16 of SEQ ID NO 86, SEQ ID NO 87, SEQ ID NO 88, SEQ ID NO 89, or SEQ ID NO 90;
b) at least one amino acid substitution relative to a wild-type equine IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 3 of SEQ ID NO: 72; and/or
c) At least one amino acid substitution relative to a wild-type equine IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 20 of SEQ ID NO: 72.
31. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising:
a) at least one amino acid substitution relative to a wild-type cat class IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises an amino acid substitution at position 16 of SEQ ID NO 86, SEQ ID NO 87, SEQ ID NO 88, SEQ ID NO 89, or SEQ ID NO 90;
b) at least one amino acid substitution relative to a wild-type equine IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises an amino acid substitution at position 3 of SEQ ID NO: 72; and/or
c) At least one amino acid substitution relative to a wild-type equine IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises an amino acid substitution at position 20 of SEQ ID NO: 72.
32. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising:
a) at least one amino acid substitution relative to a wild-type cat class IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises a proline at a position corresponding to position 16 of SEQ ID NO 86, SEQ ID NO 87, SEQ ID NO 88, SEQ ID NO 89, or SEQ ID NO 90 or at position 16 of SEQ ID NO 86, SEQ ID NO 87, SEQ ID NO 88, SEQ ID NO 89, or SEQ ID NO 90;
b) at least one amino acid substitution relative to a wild-type equine IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises a serine at a position corresponding to position 3 of SEQ ID NO:72 or at position 3 of SEQ ID NO: 72; and/or
c) At least one amino acid substitution relative to a wild-type equine IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises a proline at a position corresponding to position 20 of SEQ ID NO:72 or at position 20 of SEQ ID NO: 72.
33. The continuous polypeptide of any one of the preceding claims, comprising a variant IgG Fc polypeptide comprising a hinge region or a portion of a hinge region from an IgG Fc polypeptide of a different isotype.
34. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising a hinge region or a portion of a hinge region from a wild-type cat class IgG-1Fc polypeptide or from a wild-type equine IgG1Fc polypeptide.
35. The continuous polypeptide of any preceding claim comprising a variant IgG Fc polypeptide comprising a cysteine at a position corresponding to position 8, position 9, position 10, position 11, position 12, position 13, position 14, position 15 or position 16 of SEQ ID NO 90.
36. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising a cysteine at a position corresponding to position 14 of SEQ ID No. 90.
37. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising a cysteine at position 14 of SEQ ID No. 90.
38. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising:
a) an amino acid substitution at a position corresponding to position 21 of SEQ ID NO 34, an amino acid substitution at a position corresponding to position 23 of SEQ ID NO 34, an amino acid substitution at a position corresponding to position 25 of SEQ ID NO 34, an amino acid substitution at a position corresponding to position 80 of SEQ ID NO 34, an amino acid substitution at a position corresponding to position 205 of SEQ ID NO 34, and/or an amino acid substitution at a position corresponding to position 207 of SEQ ID NO 34;
b) an amino acid substitution at a position corresponding to position 21 of SEQ ID NO 37, an amino acid substitution at a position corresponding to position 23 of SEQ ID NO 37 and/or an amino acid substitution at a position corresponding to position 24 of SEQ ID NO 37;
c) an amino acid substitution at a position corresponding to position 21 of SEQ ID NO:39, an amino acid substitution at a position corresponding to position 23 of SEQ ID NO:39, an amino acid substitution at a position corresponding to position 25 of SEQ ID NO:39, an amino acid substitution at a position corresponding to position 80 of SEQ ID NO:39 and/or an amino acid substitution at a position corresponding to position 207 of SEQ ID NO: 39;
d) an amino acid substitution at a position corresponding to position 15 of SEQ ID NO:71 and/or an amino acid substitution at a position corresponding to position 203 of SEQ ID NO: 71;
e) an amino acid substitution at a position corresponding to position 199 of SEQ ID NO. 75 and/or an amino acid substitution at a position corresponding to position 200 of SEQ ID NO. 75; and/or
f) An amino acid substitution at a position corresponding to position 199 of SEQ ID NO:76, an amino acid substitution at a position corresponding to position 200 of SEQ ID NO:76, an amino acid substitution at a position corresponding to position 201 of SEQ ID NO:76 and/or an amino acid substitution at a position corresponding to position 202 of SEQ ID NO: 76.
39. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising:
a) an amino acid substitution at position 21 of SEQ ID NO. 34, an amino acid substitution at position 23 of SEQ ID NO. 34, an amino acid substitution at position 25 of SEQ ID NO. 34, an amino acid substitution at position 80 of SEQ ID NO. 34, an amino acid substitution at position 205 of SEQ ID NO. 34 and/or an amino acid substitution at position 207 of SEQ ID NO. 34;
b) an amino acid substitution at position 21 of SEQ ID NO 37, an amino acid substitution at position 23 of SEQ ID NO 37 and/or an amino acid substitution at position 24 of SEQ ID NO 37;
c) an amino acid substitution at position 21 of SEQ ID NO:39, an amino acid substitution at position 23 of SEQ ID NO:39, an amino acid substitution at position 25 of SEQ ID NO:39, an amino acid substitution at position 80 of SEQ ID NO:39 and/or an amino acid substitution at position 207 of SEQ ID NO: 39;
d) an amino acid substitution at position 15 of SEQ ID NO:71 and/or an amino acid substitution at position 203 of SEQ ID NO: 71;
e) an amino acid substitution at position 199 of SEQ ID NO. 75 and/or an amino acid substitution at position 200 of SEQ ID NO. 75; and/or
f) An amino acid substitution at position 199 of SEQ ID NO. 76, an amino acid substitution at position 200 of SEQ ID NO. 76, an amino acid substitution at position 201 of SEQ ID NO. 76 and/or an amino acid substitution at position 202 of SEQ ID NO. 76.
40. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising:
a) a threonine at a position corresponding to position 21 of SEQ ID NO. 34, a leucine at a position corresponding to position 23 of SEQ ID NO. 34, an alanine at a position corresponding to position 25 of SEQ ID NO. 34, a glycine at a position corresponding to position 80 of SEQ ID NO. 34, an alanine at a position corresponding to position 205 of SEQ ID NO. 34 and/or a histidine at a position corresponding to position 207 of SEQ ID NO. 34;
b) threonine at a position corresponding to position 21 of SEQ ID NO 37, leucine at a position corresponding to position 23 of SEQ ID NO 37 and/or isoleucine at a position corresponding to position 24 of SEQ ID NO 37;
c) a threonine at a position corresponding to position 21 of SEQ ID NO:39, a leucine at a position corresponding to position 23 of SEQ ID NO:39, an alanine at a position corresponding to position 25 of SEQ ID NO:39, a glycine at a position corresponding to position 80 of SEQ ID NO:39 and/or a histidine at a position corresponding to position 207 of SEQ ID NO: 39;
d) threonine or valine at a position corresponding to position 15 of SEQ ID NO:71 and/or tyrosine or valine at a position corresponding to position 203 of SEQ ID NO: 71;
e) leucine at a position corresponding to position 199 of SEQ ID No. 75 and/or histidine at a position corresponding to position 200 of SEQ ID No. 75; and/or
f) A leucine at a position corresponding to position 199 of SEQ ID NO:76, a histidine at a position corresponding to position 200 of SEQ ID NO:76, an asparagine at a position corresponding to position 201 of SEQ ID NO:76 and/or a histidine at a position corresponding to position 202 of SEQ ID NO: 76.
41. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising:
a) threonine at position 21 of SEQ ID NO. 34, leucine at position 23 of SEQ ID NO. 34, alanine at position 25 of SEQ ID NO. 34, glycine at position 80 of SEQ ID NO. 34, alanine at position 205 of SEQ ID NO. 34 and/or histidine at position 207 of SEQ ID NO. 34;
b) threonine at position 21 of SEQ ID NO 37, leucine at position 23 of SEQ ID NO 37 and/or isoleucine at position 24 of SEQ ID NO 37;
c) threonine at position 21 of SEQ ID NO:39, leucine at position 23 of SEQ ID NO:39, alanine at position 25 of SEQ ID NO:39, glycine at position 80 of SEQ ID NO:39 and/or histidine at position 207 of SEQ ID NO: 39;
d) threonine or valine at position 15 of SEQ ID NO:71 and/or tyrosine or valine at position 203 of SEQ ID NO: 71;
e) leucine at position 199 of SEQ ID NO. 75 and/or histidine at position 200 of SEQ ID NO. 75; and/or
f) Leucine at position 199 of SEQ ID NO:76, histidine at position 200 of SEQ ID NO:76, asparagine at position 201 of SEQ ID NO:76 and/or histidine at position 202 of SEQ ID NO: 76.
42. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising:
a) an amino acid substitution at a position corresponding to position 93 of SEQ ID NO 35 or an amino acid substitution at a position corresponding to position 93 of SEQ ID NO 37;
b) an amino acid substitution at a position corresponding to position 87 of SEQ ID NO 70, an amino acid substitution at a position corresponding to position 87 of SEQ ID NO 73, an amino acid substitution at a position corresponding to position 87 of SEQ ID NO 74 or an amino acid substitution at a position corresponding to position 87 of SEQ ID NO 77; or
c) An amino acid substitution at a position corresponding to position 198 of SEQ ID NO 86, an amino acid substitution at a position corresponding to position 198 of SEQ ID NO 87, an amino acid substitution at a position corresponding to position 198 of SEQ ID NO 88 or an amino acid substitution at a position corresponding to position 198 of SEQ ID NO 89.
43. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising:
a) an amino acid substitution at position 93 of SEQ ID NO 35 or an amino acid substitution at position 93 of SEQ ID NO 37;
b) an amino acid substitution at position 87 of SEQ ID NO 70, an amino acid substitution at position 87 of SEQ ID NO 73, an amino acid substitution at position 87 of SEQ ID NO 74 or an amino acid substitution at position 87 of SEQ ID NO 77; or
c) An amino acid substitution at position 198 of SEQ ID NO 86, an amino acid substitution at position 198 of SEQ ID NO 87, an amino acid substitution at position 198 of SEQ ID NO 88 or an amino acid substitution at position 198 of SEQ ID NO 89.
44. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising:
a) an arginine at a position corresponding to position 93 of SEQ ID NO 35 or an arginine at a position corresponding to position 93 of SEQ ID NO 37;
b) a serine at a position corresponding to position 87 of SEQ ID NO 70, a serine substitution at a position corresponding to position 87 of SEQ ID NO 73, a serine at a position corresponding to position 87 of SEQ ID NO 74 or a serine at a position corresponding to position 87 of SEQ ID NO 77; or
c) Alanine at a position corresponding to position 198 of SEQ ID NO 86, alanine at a position corresponding to position 198 of SEQ ID NO 87, alanine at a position corresponding to position 198 of SEQ ID NO 88 or alanine at a position corresponding to position 198 of SEQ ID NO 89.
45. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising:
a) an arginine at position 93 of SEQ ID NO 35 or an arginine at position 93 of SEQ ID NO 37;
b) serine at position 87 of SEQ ID NO. 70, serine at position 87 of SEQ ID NO. 73, serine at position 87 of SEQ ID NO. 74 or serine at position 87 of SEQ ID NO. 77; or
c) Alanine at position 198 of SEQ ID NO. 86, alanine at position 198 of SEQ ID NO. 87, alanine at position 198 of SEQ ID NO. 88 or alanine at position 198 of SEQ ID NO. 89.
46. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising:
a) (ii) tyrosine or phenylalanine at a position corresponding to position 23 of SEQ ID NO 34, 35, 37, 39, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90;
b) (ii) tyrosine at a position corresponding to position 82 of SEQ ID NO 34, 35, 37, 39, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90;
c) 35, 37, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90 and histidine at a position corresponding to position 207 thereof;
d) 35, 37, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90 and a tyrosine at a position corresponding to position 207 thereof;
e) (ii) a tyrosine at a position corresponding to position 207 of SEQ ID NO 35, SEQ ID NO 37, SEQ ID NO 70, SEQ ID NO 71, SEQ ID NO 73, SEQ ID NO 74, SEQ ID NO 75, SEQ ID NO 76, SEQ ID NO 77, SEQ ID NO 86, SEQ ID NO 88 or SEQ ID NO 90;
f) a tyrosine at a position corresponding to position 82 of SEQ ID NO:34 or SEQ ID NO:39 and a histidine at a position corresponding to position 208 thereof;
g) tyrosine at a position corresponding to position 82 of SEQ ID NO:34 or SEQ ID NO:39 and tyrosine at a position corresponding to position 208 thereof; or
h) A tyrosine at a position corresponding to position 208 of SEQ ID NO:34 or SEQ ID NO: 39.
47. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising:
a) (ii) tyrosine or phenylalanine at position 23 of SEQ ID NO 34, 35, 37, 39, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90;
b) (iii) tyrosine at position 82 of SEQ ID NO 34, 35, 37, 39, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90;
c) 35, 37, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90 and histidine at position 207;
d) 35, 37, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90 and a tyrosine at position 82 and 207;
e) (iii) tyrosine at position 207 of SEQ ID NO 35, SEQ ID NO 37, SEQ ID NO 70, SEQ ID NO 71, SEQ ID NO 73, SEQ ID NO 74, SEQ ID NO 75, SEQ ID NO 76, SEQ ID NO 77, SEQ ID NO 86, SEQ ID NO 88 or SEQ ID NO 90;
f) a tyrosine at position 82 and a histidine at position 208 of SEQ ID NO:34 or SEQ ID NO: 39;
g) tyrosine at position 82 and tyrosine at position 208 of SEQ ID NO:34 or SEQ ID NO: 39; or
h) Tyrosine at position 208 of SEQ ID NO:34 or SEQ ID NO: 39.
48. The contiguous polypeptide of any of the preceding claims, comprising a variant IgG Fc polypeptide comprising an amino acid sequence at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to an amino acid sequence of seq id no: SEQ ID NO 40, SEQ ID NO 41, SEQ ID NO 42, SEQ ID NO 43, SEQ ID NO 44, SEQ ID NO 45, SEQ ID NO 46, SEQ ID NO 47, SEQ ID NO 48, SEQ ID NO 49, SEQ ID NO 50, SEQ ID NO 51, SEQ ID NO 52, SEQ ID NO 53, SEQ ID NO 54, SEQ ID NO 55, SEQ ID NO 56, SEQ ID NO 57, SEQ ID NO 58, SEQ ID NO 59, SEQ ID NO 60, SEQ ID NO 61, SEQ ID NO 62, SEQ ID NO 63, SEQ ID NO 64, SEQ ID NO 65, SEQ ID NO 66, SEQ ID NO 67, SEQ ID NO 68, SEQ ID NO 69, SEQ ID NO 78, SEQ ID NO 79, SEQ ID NO 80, SEQ ID NO 45, SEQ ID NO 47, SEQ ID NO 48, SEQ ID NO 58, SEQ ID NO 59, SEQ ID NO 69, SEQ ID NO 78, SEQ ID NO 79, SEQ ID NO 80, SEQ ID NO, SEQ ID NO 81, SEQ ID NO 82, SEQ ID NO 83, SEQ ID NO 84, SEQ ID NO 85, SEQ ID NO 91, SEQ ID NO 92, SEQ ID NO 93, SEQ ID NO 94, SEQ ID NO 95, SEQ ID NO 96, SEQ ID NO 97, SEQ ID NO 98, SEQ ID NO 99, SEQ ID NO 100, SEQ ID NO 101, SEQ ID NO 102, SEQ ID NO 103, SEQ ID NO 104, SEQ ID NO 105, SEQ ID NO 106, SEQ ID NO 107, SEQ ID NO 108, SEQ ID NO 109, SEQ ID NO 197, SEQ ID NO 198, SEQ ID NO 199, SEQ ID NO 200, SEQ ID NO 201, SEQ ID NO 202, SEQ ID NO 203, SEQ ID NO 204, SEQ ID NO 205, SEQ ID NO, 206, 207, 208, 209 and/or 210.
49. The continuous polypeptide of any preceding claim, comprising a variant IgG Fc polypeptide comprising the amino acid sequence of: SEQ ID NO 40, SEQ ID NO 41, SEQ ID NO 42, SEQ ID NO 43, SEQ ID NO 44, SEQ ID NO 45, SEQ ID NO 46, SEQ ID NO 47, SEQ ID NO 48, SEQ ID NO 49, SEQ ID NO 50, SEQ ID NO 51, SEQ ID NO 52, SEQ ID NO 53, SEQ ID NO 54, SEQ ID NO 55, SEQ ID NO 56, SEQ ID NO 57, SEQ ID NO 58, SEQ ID NO 59, SEQ ID NO 60, SEQ ID NO 61, SEQ ID NO 62, SEQ ID NO 63, SEQ ID NO 64, SEQ ID NO 65, SEQ ID NO 66, SEQ ID NO 67, SEQ ID NO 68, SEQ ID NO 69, SEQ ID NO 78, SEQ ID NO 79, SEQ ID NO 80, SEQ ID NO 45, SEQ ID NO 47, SEQ ID NO 48, SEQ ID NO 58, SEQ ID NO 59, SEQ ID NO 69, SEQ ID NO 78, SEQ ID NO 79, SEQ ID NO 80, SEQ ID NO, SEQ ID NO 81, SEQ ID NO 82, SEQ ID NO 83, SEQ ID NO 84, SEQ ID NO 85, SEQ ID NO 91, SEQ ID NO 92, SEQ ID NO 93, SEQ ID NO 94, SEQ ID NO 95, SEQ ID NO 96, SEQ ID NO 97, SEQ ID NO 98, SEQ ID NO 99, SEQ ID NO 100, SEQ ID NO 101, SEQ ID NO 102, SEQ ID NO 103, SEQ ID NO 104, SEQ ID NO 105, SEQ ID NO 106, SEQ ID NO 107, SEQ ID NO 108, SEQ ID NO 109, SEQ ID NO 197, SEQ ID NO 198, SEQ ID NO 199, SEQ ID NO 200, SEQ ID NO 201, SEQ ID NO 202, SEQ ID NO 203, SEQ ID NO 204, SEQ ID NO 205, SEQ ID NO, 206, 207, 208, 209 and/or 210.
50. The contiguous polypeptide of any of the preceding claims, further comprising at least one extracellular domain of a NGFR polypeptide (NGFR ECD polypeptide).
51. The contiguous polypeptide of any of the preceding claims, further comprising at least one NGFR ECD polypeptide comprising an amino acid sequence of SEQ ID No. 135, SEQ ID No. 137, and/or SEQ ID No. 139.
52. The continuous polypeptide of any one of the preceding claims, comprising the amino acid sequence of: 16, 17, 18, 19, 20, 21, 22, 23, 24, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 130, 131, 132, 111, 140, 141, 142, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 186, 187, 188, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 236, 233, 234, 235, 236, 238, 239, 240, 241, 242, 243, 244, 245 or 246.
53. A contiguous polypeptide comprising the amino acid sequence of: 16, 17, 18, 19, 20, 21, 22, 23, 24, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 130, 131, 132, 111, 140, 141, 142, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 186, 187, 188, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 236, 233, 234, 235, 236, 238, 239, 240, 241, 242, 243, 244, 245 or 246.
54. The continuous polypeptide of any preceding claim, wherein the Trk A ECD polypeptide is glycosylated.
55. The continuous polypeptide of any one of the preceding claims, wherein the Trk A ECD polypeptide comprises at least one glycan moiety.
56. The continuous polypeptide of any preceding claim, wherein the Trk A ECD polypeptide is pegylated.
57. The continuous polypeptide of any preceding claim, wherein the Trk A ECD polypeptide is pegylated at a glycan, at a primary amine, and/or at an N-terminal alpha-amine.
58. An isolated nucleic acid encoding the contiguous polypeptide of any of the preceding claims.
59. A host cell comprising the nucleic acid of claim 58.
60. A method of producing a polypeptide, the method comprising culturing the host cell of claim 59 and isolating the contiguous polypeptide.
61. A pharmaceutical composition comprising the continuous polypeptide of any one of claims 1 to 57 and a pharmaceutically acceptable carrier.
62. The pharmaceutical composition according to claim 61, wherein the pharmaceutically acceptable carrier comprises about 5 to about 50mM sodium citrate; about 5 to about 50mM histidine; or about 5 to about 50mM sodium acetate.
63. The pharmaceutical composition of claim 61 or claim 62, wherein the pharmaceutical composition has a pH of 5 to 6.
64. A method of treating a companion animal species suffering from an NGF-induced disorder, comprising administering to the companion animal species a therapeutically effective amount of the continuous polypeptide of any one of claims 1 to 57 or the pharmaceutical composition of any one of claims 61 to 63.
65. A method of treating a companion animal species suffering from pain, the method comprising administering to the companion animal species a therapeutically effective amount of the continuous polypeptide of any one of claims 1-57 or the pharmaceutical composition of any one of claims 61-63.
66. The method of claim 64 or claim 65 wherein the companion animal species is canine, feline, or equine.
67. The method according to any one of claims 64 to 66, wherein the NGF-induced condition or the pain is chronic pain, acute pain and/or inflammatory pain.
68. The method according to any one of claims 64 to 67, wherein the NGF-induced condition or the pain is osteoarthritis pain, back pain, cancer pain and/or neuropathic pain.
69. The method of any one of claims 64 to 68, wherein the NGF-induced condition or the pain is pain associated with surgery, a broken bone or fracture, dental treatment, a burn, a cut, and/or childbirth.
70. The method of any one of claims 64-69, wherein the continuous polypeptide or the pharmaceutical composition is administered parenterally.
71. The method of any one of claims 64-70, wherein said continuous polypeptide or said pharmaceutical composition is administered by an intramuscular route, an intraperitoneal route, an intracerobrospinal route, a subcutaneous route, an intraarterial route, an intrasynovial route, an intrathecal route, or an inhalation route.
72. The method of any one of claims 64 to 71, wherein the method further comprises administering an NGF kinase inhibitor, a PI3K inhibitor, a ras inhibitor, a CGRP inhibitor, a TNF inhibitor, an IL17 inhibitor, an EGFR inhibitor, and/or a phospholipase C pathway inhibitor.
73. The method of any one of claims 64-72, wherein the method further comprises administering one or more pain treatment drugs, such as corticosteroids, non-steroidal anti-inflammatory drugs (NSAIDs), cyclooxygenase inhibitors, opioids, and/or cannabinoids.
74. A method of reducing NGF signaling activity in a cell, the method comprising exposing the cell to the continuous polypeptide of any one of claims 1 to 57 or the pharmaceutical composition of any one of claims 61 to 63 under conditions wherein: the conditions allow binding of the contiguous polypeptide to NGF.
75. The method of claim 74, wherein the cells are exposed to the continuous polypeptide or the pharmaceutical composition ex vivo.
76. The method of claim 74, wherein the cell is exposed to the continuous polypeptide or the pharmaceutical composition in vivo.
77. The method of any one of claims 74-76, wherein the cell is a canine cell, a feline cell, or an equine cell.
78. A method for detecting NGF in a sample from a companion animal species, the method comprising contacting the sample with the contiguous polypeptide of any one of claims 1 to 57 or the pharmaceutical composition of any one of claims 61 to 63 under conditions wherein: the conditions allow binding of the contiguous polypeptide to NGF; and detecting whether a complex is formed between the polypeptide and NGF in the sample.
79. The method of claim 78, wherein the sample is a biological sample obtained from a canine, a feline, or an equine.
Technical Field
The present disclosure relates to NGF-binding polypeptides comprising the extracellular domain of TrkA from a companion animal species. The disclosure also relates to methods of using the polypeptides, e.g., for treating NGF-induced disorders or reducing NGF signaling activity in cells, e.g., in companion animals such as canines, felines, and equines.
Background
Nerve Growth Factor (NGF) is a neurotrophic factor that has a broad effect on regulating the growth, maintenance, proliferation and survival of certain neurons. NGF is also associated with chronic pain and inflammatory pain. NGF binds to two types of receptors: tropomyosin (tropimyosine) receptor kinase a (trka) and low affinity NGF receptors. When NGF (a dimer) binds to the TrkA extracellular domain, it causes receptor dimerization, thereby activating downstream kinase activity. The TrkA extracellular domain may be used to antagonize NGF activity, reduce free NGF, and/or attenuate clinical signs and symptoms associated with NGF-related pain.
Companion species animals (e.g., cats, dogs, and horses) suffer from a number of conditions similar to those of humans, including chronic pain and inflammatory pain. Thus, there remains a need for methods and species-specific compounds that can be used specifically to bind NGF in companion animals to treat NGF-induced disorders and reduce NGF signaling activity.
Disclosure of Invention
Embodiment 1. a continuous polypeptide comprising at least one extracellular domain of a TrkA polypeptide (TrkA ECD polypeptide) from a companion animal species and a fusion partner.
Embodiment 2. the continuous polypeptide of embodiment 1, wherein the continuous polypeptide is less than 5x10 as measured by biofilm layer interferometry-6M, less than 1x10-6M, less than 5x10-7M, less than 1x10-7M, less than 5x10-8M, less than 1x10-8M, less than 5x10-9M, less than 1x10-9M, less than 5x10-10M, less than 1x10-10M, less than 5x10-11M, less than 1x10-11M, less than 5x10-12M or less than 1x10-12The dissociation constant (Kd) of M binds to the NGF polypeptide.
Embodiment 3. the continuous polypeptide of embodiment 2, wherein the NGF polypeptide is a human NGF polypeptide, a canine NGF polypeptide, a feline NGF polypeptide, or an equine NGF polypeptide.
Embodiment 4. the continuous polypeptide of any one of the preceding embodiments, wherein the continuous polypeptide reduces NGF signaling in the companion animal species.
Embodiment 5. the continuous polypeptide of any one of the preceding embodiments, wherein the companion animal species is canine, feline, or equine.
Embodiment 6. the contiguous polypeptide of any of the preceding embodiments, wherein the amino acid sequence of the TrkA ECD polypeptide is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence of seq id no: SEQ ID NO 2, SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 5, SEQ ID NO 7, SEQ ID NO 8, SEQ ID NO 9, SEQ ID NO 10, SEQ ID NO 12, SEQ ID NO 13, SEQ ID NO 14 or SEQ ID NO 15.
Embodiment 7. the continuous polypeptide of any one of the preceding embodiments, wherein the TrkA ECD polypeptide comprises:
a) a cysteine at a position corresponding to position 7 and position 89 of SEQ ID NO 2,3, 4, 7, 8, 9, 12, 13 or 14; or
b) A cysteine at a position corresponding to position 5 and position 87 of SEQ ID NO 5, SEQ ID NO 10 or SEQ ID NO 15.
Embodiment 8 the continuous polypeptide of any one of the preceding embodiments, wherein the TrkA ECD polypeptide comprises:
a) a cysteine at position 7 and position 89 of SEQ ID NO 2,3, 4, 7, 8, 9, 12, 13 or 14; or
b) Cysteines at positions 5 and 87 of SEQ ID NO 5, 10 or 15.
Embodiment 9 the contiguous polypeptide of any one of the preceding embodiments, wherein the TrkA ECD polypeptide comprises at least one N-linked glycosylation site that is not present in a corresponding wild-type TrkA ECD polypeptide, wherein the N-linked glycosylation site comprises the sequence asparagine-xaa-serine or asparagine-xaa-threonine, wherein xaa is any amino acid other than proline, and wherein one N-linked glycosylation site does not overlap with another N-linked glycosylation site.
Embodiment 10 the continuous polypeptide of any one of the preceding embodiments, wherein the TrkA ECD comprises at least one N-linked glycosylation site at one or more positions selected from the group consisting of:
a) 3, 4, 8, 9, 13 or 14 amino acid positions 6-8, 31-33, 84-86, 85-87, 86-88, 88-90, 90-92, 92-94 and/or 94-96; or
b) Amino acid positions 4-6, 29-31, 82-84, 83-85, 84-86, 86-88, 89-90, 90-92 and/or 92-94 of SEQ ID NO 5, SEQ ID NO 10 or SEQ ID NO 15.
Embodiment 11 the continuous polypeptide of any one of the preceding embodiments, wherein the TrkA ECD polypeptide comprises:
a) an amino acid other than proline at an amino acid position corresponding to position 30 and/or position 85 of SEQ ID NO 3, 4, 8, 9, 13 or 14; and/or
b) An amino acid other than proline at an amino acid position corresponding to position 28 or position 83 of SEQ ID NO 5, SEQ ID NO 10 or SEQ ID NO 15.
Embodiment 12. the continuous polypeptide of any one of the preceding embodiments, wherein the TrkA ECD polypeptide comprises:
a) valine, glutamic acid, alanine or isoleucine at an amino acid position corresponding to position 30 and/or position 85 of SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 8, SEQ ID NO 9, SEQ ID NO 13 or SEQ ID NO 14; and/or
b) Valine, glutamic acid, alanine or isoleucine at an amino acid position corresponding to position 28 or 83 of SEQ ID NO 5, 10 or 15.
Embodiment 13. the continuous polypeptide of any one of the preceding embodiments, wherein the TrkA ECD polypeptide comprises one or more amino acid modifications listed in table a:
wherein X indicates any amino acid other than proline (e.g. E, V, A or I);
table B:
wherein X indicates any amino acid other than proline (e.g. E, V, A or I); and/or
Table C:
wherein X indicates any amino acid other than proline (e.g. E, V, A or I).
Embodiment 14. the continuous polypeptide of any one of the preceding embodiments, wherein the TrkA ECD polypeptide comprises an amino acid sequence selected from the group consisting of: SEQ ID NO 2, SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 5, SEQ ID NO 7, SEQ ID NO 8, SEQ ID NO 9, SEQ ID NO 10, SEQ ID NO 12, SEQ ID NO 13, SEQ ID NO 14, SEQ ID NO 15, SEQ ID NO 25, SEQ ID NO 26, SEQ ID NO 27, SEQ ID NO 28, SEQ ID NO 29, SEQ ID NO 30, SEQ ID NO 31, SEQ ID NO 32 and SEQ ID NO 33.
Embodiment 15. the continuous polypeptide of any one of the preceding embodiments, wherein the continuous polypeptide comprises a linker.
Embodiment 16. the continuous polypeptide of embodiment 15, wherein the linker comprises an amino acid sequence selected from the group consisting of: G. GG, GGG, S, SS, SSS, GS, GSGS (SEQ ID NO:143), GSGSGS (SEQ ID NO:144), GGS, GGSGGS (SEQ ID NO:145), GGSGGSGGS (SEQ ID NO:146), GGGS (SEQ ID NO:147), GGGSGGGS (SEQ ID NO:148), GGGSGGGSGGGS (SEQ ID NO:149), GSS, GSSGSS (SEQ ID NO:150), GSSGSSGSS (SEQ ID NO:151), GGSS (SEQ ID NO:152), GGSSGGSS (SEQ ID NO:153), GGSSGGSSGGS (SEQ ID NO:154), SGGSGGS (SEQ ID NO:155) and SGGGSGGGS (SEQ ID NO: 156).
Embodiment 17. the continuous polypeptide of any one of the preceding embodiments, wherein the fusion partner is selected from the group consisting of an Fc polypeptide, albumin, and an albumin binding fragment.
Embodiment 18. the continuous polypeptide of any one of the preceding embodiments, wherein the fusion partner is an Fc polypeptide comprising (a) a wild-type or variant canine IgG-A, IgG-B, IgG-C or IgG-D polypeptide; (b) a wild-type or variant cat class IgG1a, IgG1b, or IgG2 polypeptide; or (c) a wild-type or variant equine IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, or IgG7 polypeptide.
Embodiment 19. the continuous polypeptide according to any one of the preceding embodiments, comprising:
formula (I): TrkA ECD 1-L1-Fc;
formula (II): Fc-L1-TrkA ECD 1;
formula (III): TrkA ECD 1-L1-Fc-L2-TrkA ECD 2;
formula (IV): TrkA ECD 1-L1-TrkA ECD 2-L2-Fc; or
Formula (V): Fc-L1-TrkA ECD 1-L2-TrkA ECD 2,
wherein TrkA ECD 1 is a first TrkA ECD polypeptide, TrkA ECD 2 is a second TrkA ECD polypeptide, L1 and L2 are optional linkers, and Fc is a wild-type or variant IgG Fc polypeptide of an accompanying animal species.
Embodiment 20 the continuous polypeptide of embodiment 19, wherein TrkA ECD 1 and TrkA ECD 2 are the same polypeptide.
Embodiment 21. the contiguous polypeptide of embodiment 19, wherein TrkA ECD 1 and TrkA ECD 2 are different polypeptides.
Embodiment 22. the continuous polypeptide of any one of the preceding embodiments, wherein the fusion partner or Fc is a variant Fc polypeptide comprising:
a) at least one amino acid modification relative to a wild-type IgG Fc polypeptide of a companion animal species, wherein the variant IgG Fc polypeptide has increased binding affinity for protein a relative to the wild-type IgG Fc polypeptide;
b) at least one amino acid modification relative to a wild-type IgG Fc polypeptide of a companion animal species, wherein the variant IgG Fc polypeptide has reduced binding affinity to C1q relative to the wild-type IgG Fc polypeptide;
c) at least one amino acid modification relative to a wild-type IgG Fc polypeptide of a companion animal species, wherein the variant IgG Fc polypeptide has reduced binding affinity to CD16 relative to the wild-type IgG Fc polypeptide;
d) a hinge region comprising at least one amino acid modification relative to a wild-type feline or equine IgG Fc polypeptide;
e) at least one amino acid substitution relative to a wild-type cat class IgG Fc polypeptide, wherein the at least one amino acid substitution is cysteine, and wherein the variant IgG Fc polypeptide is capable of forming at least one additional interchain disulfide bond relative to the wild-type cat class IgG Fc polypeptide; and/or
f) At least one amino acid substitution relative to a wild-type IgG Fc polypeptide derived from a companion animal species, wherein the variant Fc polypeptide is capable of binding to a neonatal Fc receptor (FcRn) with increased affinity relative to the wild-type Fc polypeptide.
Embodiment 23. the continuous polypeptide of any of the preceding embodiments, comprising a variant IgG Fc polypeptide that is greater than 5x10 as measured by biofilm layer interferometry-6M, greater than 1x10-5M, greater than 5x10-5M, greater than 1x10-4M, greater than 5x10-4M or greater than 1x10-3The dissociation constant (Kd) of M binds to C1q and/or CD 16.
Embodiment 24. the continuous polypeptide of any of the preceding embodiments, comprising a variant IgG Fc polypeptide that is less than 5x10 as measured by biofilm layer interferometry-6M, less than 1x10-6M, less than 5x10-7M, less than 1x10-7M, less than 5x10-8M, less than 1x10-8M, less than 5x10-9M, less than 1x10-9M, less than 5x10-10M, less than 1x10-10M, less than 5x10-11M, less than 1x10-11M, less than 5x10-12M or less than 1x10-12The dissociation constant (Kd) of M binds to protein A.
The continuous polypeptide of any of the preceding claims comprising a variant IgG Fc polypeptide that binds FcRn with greater affinity than the wild-type IgG Fc polypeptide as measured by biofilm layer interference, surface plasmon resonance, or any protein-protein interaction means at a pH in the range of about 5.0 to about 6.5, as measured at a pH of about 5.0, a pH of about 5.2, a pH of about 5.5, a pH of about 6.0, a pH of about 6.2, or a pH of about 6.5.
The continuous polypeptide of any of the preceding claims, comprising a variant IgG Fc polypeptide that is less than 5x10-6M, less than 1x10-6M, less than 5x10-7M, less than 1x10- 7M, less than 5x10-8M, less than 1x10-8M, less than 5x10-9M, less than 1x10-9M, less than 5x10-10M, less than 1x10-10M, less than 5x10-11M, less than 1x10-11M, less than 5x10-12M or less than 1x10-12The dissociation constant (Kd) of M binds to FcRn, as measured by biofilm layer interferometry, surface plasmon resonance, or any protein-protein interaction means at a pH in the range of about 5.0 to about 6.5, such as at a pH of about 5.0, a pH of about 5.5, a pH of about 6.0, or a pH of about 6.5.
The contiguous polypeptide of any of the preceding claims, comprising a variant IgG Fc polypeptide that binds to FcRn with increased affinity relative to the wild-type Fc polypeptide, and wherein the contiguous polypeptide has increased serum half-life relative to a contiguous polypeptide comprising a wild-type Fc polypeptide.
Embodiment 28. the continuous polypeptide of any one of the preceding embodiments, wherein the wild-type IgG Fc polypeptide is:
a) a canine IgG-A Fc, IgG-B Fc, IgG-C Fc, or IgG-D Fc;
b) equine IgG1Fc, IgG2Fc, IgG3 Fc, IgG4 Fc, IgG5 Fc, IgG6Fc, or IgG7 Fc; or
c) Cat class IgG1a Fc, IgG1b Fc or IgG2 Fc.
Embodiment 29. the continuous polypeptide of any one of the preceding embodiments, wherein the wild-type IgG Fc polypeptide comprises the amino acid sequence of: SEQ ID NO 34, 35, 36, 37, 38, 39, 70, 71, 72, 73, 74, 75, 76, 77, 86, 87, 88, 89 or 90.
Embodiment 30. the continuous polypeptide of any one of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising:
a) at least one amino acid substitution relative to a wild-type cat class IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 16 of SEQ ID NO 86, SEQ ID NO 87, SEQ ID NO 88, SEQ ID NO 89, or SEQ ID NO 90;
b) at least one amino acid substitution relative to a wild-type equine IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 3 of SEQ ID NO: 72; and/or
c) At least one amino acid substitution relative to a wild-type equine IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 20 of SEQ ID NO: 72.
The continuous polypeptide of any of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising:
a) at least one amino acid substitution relative to a wild-type cat class IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises an amino acid substitution at position 16 of SEQ ID NO 86, SEQ ID NO 87, SEQ ID NO 88, SEQ ID NO 89, or SEQ ID NO 90;
b) at least one amino acid substitution relative to a wild-type equine IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises an amino acid substitution at position 3 of SEQ ID NO: 72; and/or
c) At least one amino acid substitution relative to a wild-type equine IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises an amino acid substitution at position 20 of SEQ ID NO: 72.
Embodiment 32. the continuous polypeptide of any one of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising:
a) at least one amino acid substitution relative to a wild-type cat class IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises a proline at a position corresponding to position 16 of SEQ ID NO 86, SEQ ID NO 87, SEQ ID NO 88, SEQ ID NO 89, or SEQ ID NO 90 or at position 16 of SEQ ID NO 86, SEQ ID NO 87, SEQ ID NO 88, SEQ ID NO 89, or SEQ ID NO 90;
b) at least one amino acid substitution relative to a wild-type equine IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises a serine at a position corresponding to position 3 of SEQ ID NO:72 or at position 3 of SEQ ID NO: 72; and/or
c) At least one amino acid substitution relative to a wild-type equine IgG Fc polypeptide, wherein the variant IgG Fc polypeptide comprises a proline at a position corresponding to position 20 of SEQ ID NO:72 or at position 20 of SEQ ID NO: 72.
Embodiment 33. the continuous polypeptide of any one of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising a hinge region or a portion of a hinge region from an IgG Fc polypeptide of a different isotype.
The continuous polypeptide of any of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising a hinge region or a portion of a hinge region from a wild-type cat class IgG-1Fc polypeptide or from a wild-type equine IgG1Fc polypeptide.
Embodiment 35. the continuous polypeptide according to any one of the preceding embodiments, comprising a variant IgG Fc-polypeptide comprising a cysteine at a position corresponding to position 8, position 9, position 10, position 11, position 12, position 13, position 14, position 15 or position 16 of SEQ ID No. 90.
Embodiment 36. the continuous polypeptide of any one of the preceding embodiments, comprising a variant IgG Fc-polypeptide comprising a cysteine at a position corresponding to position 14 of SEQ ID NO: 90.
Embodiment 37. the continuous polypeptide of any one of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising a cysteine at position 14 of SEQ ID NO: 90.
The continuous polypeptide of any of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising:
a) an amino acid substitution at a position corresponding to position 21 of SEQ ID NO 34, an amino acid substitution at a position corresponding to position 23 of SEQ ID NO 34, an amino acid substitution at a position corresponding to position 25 of SEQ ID NO 34, an amino acid substitution at a position corresponding to position 80 of SEQ ID NO 34, an amino acid substitution at a position corresponding to position 205 of SEQ ID NO 34, and/or an amino acid substitution at a position corresponding to position 207 of SEQ ID NO 34;
b) an amino acid substitution at a position corresponding to position 21 of SEQ ID NO 37, an amino acid substitution at a position corresponding to position 23 of SEQ ID NO 37 and/or an amino acid substitution at a position corresponding to position 24 of SEQ ID NO 37;
c) an amino acid substitution at a position corresponding to position 21 of SEQ ID NO:39, an amino acid substitution at a position corresponding to position 23 of SEQ ID NO:39, an amino acid substitution at a position corresponding to position 25 of SEQ ID NO:39, an amino acid substitution at a position corresponding to position 80 of SEQ ID NO:39 and/or an amino acid substitution at a position corresponding to position 207 of SEQ ID NO: 39;
d) an amino acid substitution at a position corresponding to position 15 of SEQ ID NO:71 and/or an amino acid substitution at a position corresponding to position 203 of SEQ ID NO: 71;
e) an amino acid substitution at a position corresponding to position 199 of SEQ ID NO. 75 and/or an amino acid substitution at a position corresponding to position 200 of SEQ ID NO. 75; and/or
f) An amino acid substitution at a position corresponding to position 199 of SEQ ID NO:76, an amino acid substitution at a position corresponding to position 200 of SEQ ID NO:76, an amino acid substitution at a position corresponding to position 201 of SEQ ID NO:76 and/or an amino acid substitution at a position corresponding to position 202 of SEQ ID NO: 76.
Embodiment 39 the continuous polypeptide of any one of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising:
a) an amino acid substitution at position 21 of SEQ ID NO. 34, an amino acid substitution at position 23 of SEQ ID NO. 34, an amino acid substitution at position 25 of SEQ ID NO. 34, an amino acid substitution at position 80 of SEQ ID NO. 34, an amino acid substitution at position 205 of SEQ ID NO. 34 and/or an amino acid substitution at position 207 of SEQ ID NO. 34;
b) an amino acid substitution at position 21 of SEQ ID NO 37, an amino acid substitution at position 23 of SEQ ID NO 37 and/or an amino acid substitution at position 24 of SEQ ID NO 37;
c) an amino acid substitution at position 21 of SEQ ID NO:39, an amino acid substitution at position 23 of SEQ ID NO:39, an amino acid substitution at position 25 of SEQ ID NO:39, an amino acid substitution at position 80 of SEQ ID NO:39 and/or an amino acid substitution at position 207 of SEQ ID NO: 39;
d) an amino acid substitution at position 15 of SEQ ID NO:71 and/or an amino acid substitution at position 203 of SEQ ID NO: 71;
e) an amino acid substitution at position 199 of SEQ ID NO. 75 and/or an amino acid substitution at position 200 of SEQ ID NO. 75; and/or
f) An amino acid substitution at position 199 of SEQ ID NO. 76, an amino acid substitution at position 200 of SEQ ID NO. 76, an amino acid substitution at position 201 of SEQ ID NO. 76 and/or an amino acid substitution at position 202 of SEQ ID NO. 76.
Embodiment 40. the continuous polypeptide of any one of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising:
a) a threonine at a position corresponding to position 21 of SEQ ID NO. 34, a leucine at a position corresponding to position 23 of SEQ ID NO. 34, an alanine at a position corresponding to position 25 of SEQ ID NO. 34, a glycine at a position corresponding to position 80 of SEQ ID NO. 34, an alanine at a position corresponding to position 205 of SEQ ID NO. 34 and/or a histidine at a position corresponding to position 207 of SEQ ID NO. 34;
b) threonine at a position corresponding to position 21 of SEQ ID NO 37, leucine at a position corresponding to position 23 of SEQ ID NO 37 and/or isoleucine at a position corresponding to position 24 of SEQ ID NO 37;
c) a threonine at a position corresponding to position 21 of SEQ ID NO:39, a leucine at a position corresponding to position 23 of SEQ ID NO:39, an alanine at a position corresponding to position 25 of SEQ ID NO:39, a glycine at a position corresponding to position 80 of SEQ ID NO:39 and/or a histidine at a position corresponding to position 207 of SEQ ID NO: 39;
d) threonine or valine at a position corresponding to position 15 of SEQ ID NO:71 and/or tyrosine or valine at a position corresponding to position 203 of SEQ ID NO: 71;
e) leucine at a position corresponding to position 199 of SEQ ID No. 75 and/or histidine at a position corresponding to position 200 of SEQ ID No. 75; and/or
f) A leucine at a position corresponding to position 199 of SEQ ID NO:76, a histidine at a position corresponding to position 200 of SEQ ID NO:76, an asparagine at a position corresponding to position 201 of SEQ ID NO:76 and/or a histidine at a position corresponding to position 202 of SEQ ID NO: 76.
The continuous polypeptide of any of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising:
a) threonine at position 21 of SEQ ID NO. 34, leucine at position 23 of SEQ ID NO. 34, alanine at position 25 of SEQ ID NO. 34, glycine at position 80 of SEQ ID NO. 34, alanine at position 205 of SEQ ID NO. 34 and/or histidine at position 207 of SEQ ID NO. 34;
b) threonine at position 21 of SEQ ID NO 37, leucine at position 23 of SEQ ID NO 37 and/or isoleucine at position 24 of SEQ ID NO 37;
c) threonine at position 21 of SEQ ID NO:39, leucine at position 23 of SEQ ID NO:39, alanine at position 25 of SEQ ID NO:39, glycine at position 80 of SEQ ID NO:39 and/or histidine at position 207 of SEQ ID NO: 39;
d) threonine or valine at position 15 of SEQ ID NO:71 and/or tyrosine or valine at position 203 of SEQ ID NO: 71;
e) leucine at position 199 of SEQ ID NO. 75 and/or histidine at position 200 of SEQ ID NO. 75; and/or
f) Leucine at position 199 of SEQ ID NO:76, histidine at position 200 of SEQ ID NO:76, asparagine at position 201 of SEQ ID NO:76 and/or histidine at position 202 of SEQ ID NO: 76.
The continuous polypeptide of any of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising:
a) an amino acid substitution at a position corresponding to position 93 of SEQ ID NO 35 or an amino acid substitution at a position corresponding to position 93 of SEQ ID NO 37;
b) an amino acid substitution at a position corresponding to position 87 of SEQ ID NO 70, an amino acid substitution at a position corresponding to position 87 of SEQ ID NO 73, an amino acid substitution at a position corresponding to position 87 of SEQ ID NO 74 or an amino acid substitution at a position corresponding to position 87 of SEQ ID NO 77; or
c) An amino acid substitution at a position corresponding to position 198 of SEQ ID NO 86, an amino acid substitution at a position corresponding to position 198 of SEQ ID NO 87, an amino acid substitution at a position corresponding to position 198 of SEQ ID NO 88 or an amino acid substitution at a position corresponding to position 198 of SEQ ID NO 89.
Embodiment 43. the continuous polypeptide of any one of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising:
a) an amino acid substitution at position 93 of SEQ ID NO 35 or an amino acid substitution at position 93 of SEQ ID NO 37;
b) an amino acid substitution at position 87 of SEQ ID NO 70, an amino acid substitution at position 87 of SEQ ID NO 73, an amino acid substitution at position 87 of SEQ ID NO 74 or an amino acid substitution at position 87 of SEQ ID NO 77; or
c) An amino acid substitution at position 198 of SEQ ID NO 86, an amino acid substitution at position 198 of SEQ ID NO 87, an amino acid substitution at position 198 of SEQ ID NO 88 or an amino acid substitution at position 198 of SEQ ID NO 89.
The continuous polypeptide of any of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising:
a) an arginine at a position corresponding to position 93 of SEQ ID NO 35 or an arginine at a position corresponding to position 93 of SEQ ID NO 37;
b) a serine at a position corresponding to position 87 of SEQ ID NO 70, a serine substitution at a position corresponding to position 87 of SEQ ID NO 73, a serine at a position corresponding to position 87 of SEQ ID NO 74 or a serine at a position corresponding to position 87 of SEQ ID NO 77; or
c) Alanine at a position corresponding to position 198 of SEQ ID NO 86, alanine at a position corresponding to position 198 of SEQ ID NO 87, alanine at a position corresponding to position 198 of SEQ ID NO 88 or alanine at a position corresponding to position 198 of SEQ ID NO 89.
Embodiment 45. the continuous polypeptide of any one of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising:
a) an arginine at position 93 of SEQ ID NO 35 or an arginine at position 93 of SEQ ID NO 37;
b) serine at position 87 of SEQ ID NO. 70, serine at position 87 of SEQ ID NO. 73, serine at position 87 of SEQ ID NO. 74 or serine at position 87 of SEQ ID NO. 77; or
c) Alanine at position 198 of SEQ ID NO. 86, alanine at position 198 of SEQ ID NO. 87, alanine at position 198 of SEQ ID NO. 88 or alanine at position 198 of SEQ ID NO. 89.
Embodiment 46. the continuous polypeptide of any one of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising:
a) (ii) tyrosine or phenylalanine at a position corresponding to position 23 of SEQ ID NO 34, 35, 37, 39, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90;
b) (ii) tyrosine at a position corresponding to position 82 of SEQ ID NO 34, 35, 37, 39, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90;
c) 35, 37, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90 and histidine at a position corresponding to position 207 thereof;
d) 35, 37, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90 and a tyrosine at a position corresponding to position 207 thereof;
e) (ii) a tyrosine at a position corresponding to position 207 of SEQ ID NO 35, SEQ ID NO 37, SEQ ID NO 70, SEQ ID NO 71, SEQ ID NO 73, SEQ ID NO 74, SEQ ID NO 75, SEQ ID NO 76, SEQ ID NO 77, SEQ ID NO 86, SEQ ID NO 88 or SEQ ID NO 90;
f) a tyrosine at a position corresponding to position 82 of SEQ ID NO:34 or SEQ ID NO:39 and a histidine at a position corresponding to position 208 thereof;
g) tyrosine at a position corresponding to position 82 of SEQ ID NO:34 or SEQ ID NO:39 and tyrosine at a position corresponding to position 208 thereof; or
h) A tyrosine at a position corresponding to position 208 of SEQ ID NO:34 or SEQ ID NO: 39.
The continuous polypeptide of any of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising:
a) (ii) tyrosine or phenylalanine at position 23 of SEQ ID NO 34, 35, 37, 39, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90;
b) (iii) tyrosine at position 82 of SEQ ID NO 34, 35, 37, 39, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90;
c) 35, 37, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90 and histidine at position 207;
d) 35, 37, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90 and a tyrosine at position 82 and 207;
e) (iii) tyrosine at position 207 of SEQ ID NO 35, SEQ ID NO 37, SEQ ID NO 70, SEQ ID NO 71, SEQ ID NO 73, SEQ ID NO 74, SEQ ID NO 75, SEQ ID NO 76, SEQ ID NO 77, SEQ ID NO 86, SEQ ID NO 88 or SEQ ID NO 90;
f) a tyrosine at position 82 and a histidine at position 208 of SEQ ID NO:34 or SEQ ID NO: 39;
g) tyrosine at position 82 and tyrosine at position 208 of SEQ ID NO:34 or SEQ ID NO: 39; or
h) Tyrosine at position 208 of SEQ ID NO:34 or SEQ ID NO: 39.
Embodiment 48 the continuous polypeptide of any one of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising an amino acid sequence at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence of seq id no: SEQ ID NO 40, SEQ ID NO 41, SEQ ID NO 42, SEQ ID NO 43, SEQ ID NO 44, SEQ ID NO 45, SEQ ID NO 46, SEQ ID NO 47, SEQ ID NO 48, SEQ ID NO 49, SEQ ID NO 50, SEQ ID NO 51, SEQ ID NO 52, SEQ ID NO 53, SEQ ID NO 54, SEQ ID NO 55, SEQ ID NO 56, SEQ ID NO 57, SEQ ID NO 58, SEQ ID NO 59, SEQ ID NO 60, SEQ ID NO 61, SEQ ID NO 62, SEQ ID NO 63, SEQ ID NO 64, SEQ ID NO 65, SEQ ID NO 66, SEQ ID NO 67, SEQ ID NO 68, SEQ ID NO 69, SEQ ID NO 78, SEQ ID NO 79, SEQ ID NO 80, SEQ ID NO 45, SEQ ID NO 47, SEQ ID NO 48, SEQ ID NO 58, SEQ ID NO 59, SEQ ID NO 69, SEQ ID NO 78, SEQ ID NO 79, SEQ ID NO 80, SEQ ID NO, SEQ ID NO 81, SEQ ID NO 82, SEQ ID NO 83, SEQ ID NO 84, SEQ ID NO 85, SEQ ID NO 91, SEQ ID NO 92, SEQ ID NO 93, SEQ ID NO 94, SEQ ID NO 95, SEQ ID NO 96, SEQ ID NO 97, SEQ ID NO 98, SEQ ID NO 99, SEQ ID NO 100, SEQ ID NO 101, SEQ ID NO 102, SEQ ID NO 103, SEQ ID NO 104, SEQ ID NO 105, SEQ ID NO 106, SEQ ID NO 107, SEQ ID NO 108, SEQ ID NO 109, SEQ ID NO 197, SEQ ID NO 198, SEQ ID NO 199, SEQ ID NO 200, SEQ ID NO 201, SEQ ID NO 202, SEQ ID NO 203, SEQ ID NO 204, SEQ ID NO 205, SEQ ID NO, 206, 207, 208, 209 and/or 210.
Embodiment 49. the continuous polypeptide of any one of the preceding embodiments, comprising a variant IgG Fc polypeptide comprising the amino acid sequence of: SEQ ID NO 40, SEQ ID NO 41, SEQ ID NO 42, SEQ ID NO 43, SEQ ID NO 44, SEQ ID NO 45, SEQ ID NO 46, SEQ ID NO 47, SEQ ID NO 48, SEQ ID NO 49, SEQ ID NO 50, SEQ ID NO 51, SEQ ID NO 52, SEQ ID NO 53, SEQ ID NO 54, SEQ ID NO 55, SEQ ID NO 56, SEQ ID NO 57, SEQ ID NO 58, SEQ ID NO 59, SEQ ID NO 60, SEQ ID NO 61, SEQ ID NO 62, SEQ ID NO 63, SEQ ID NO 64, SEQ ID NO 65, SEQ ID NO 66, SEQ ID NO 67, SEQ ID NO 68, SEQ ID NO 69, SEQ ID NO 78, SEQ ID NO 79, SEQ ID NO 80, SEQ ID NO 45, SEQ ID NO 47, SEQ ID NO 48, SEQ ID NO 58, SEQ ID NO 59, SEQ ID NO 69, SEQ ID NO 78, SEQ ID NO 79, SEQ ID NO 80, SEQ ID NO, SEQ ID NO 81, SEQ ID NO 82, SEQ ID NO 83, SEQ ID NO 84, SEQ ID NO 85, SEQ ID NO 91, SEQ ID NO 92, SEQ ID NO 93, SEQ ID NO 94, SEQ ID NO 95, SEQ ID NO 96, SEQ ID NO 97, SEQ ID NO 98, SEQ ID NO 99, SEQ ID NO 100, SEQ ID NO 101, SEQ ID NO 102, SEQ ID NO 103, SEQ ID NO 104, SEQ ID NO 105, SEQ ID NO 106, SEQ ID NO 107, SEQ ID NO 108, SEQ ID NO 109, SEQ ID NO 197, SEQ ID NO 198, SEQ ID NO 199, SEQ ID NO 200, SEQ ID NO 201, SEQ ID NO 202, SEQ ID NO 203, SEQ ID NO 204, SEQ ID NO 205, SEQ ID NO, 206, 207, 208, 209 and/or 210.
Embodiment 50. the contiguous polypeptide according to any of the preceding embodiments, further comprising at least one extracellular domain of a NGFR polypeptide (NGFR ECD polypeptide).
Embodiment 51. the contiguous polypeptide according to any of the preceding embodiments, further comprising at least one NGFR ECD polypeptide comprising the amino acid sequence of SEQ ID NO:135, SEQ ID NO:137 and/or SEQ ID NO: 139.
Embodiment 52. the continuous polypeptide according to any one of the preceding embodiments, comprising the amino acid sequence of: 16, 17, 18, 19, 20, 21, 22, 23, 24, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 130, 131, 132, 111, 140, 141, 142, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 186, 187, 188, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 236, 233, 234, 235, 236, 238, 239, 240, 241, 242, 243, 244, 245 or 246.
Embodiment 53. a continuous polypeptide comprising the amino acid sequence of: 16, 17, 18, 19, 20, 21, 22, 23, 24, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 130, 131, 132, 111, 140, 141, 142, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 186, 187, 188, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 236, 233, 234, 235, 236, 238, 239, 240, 241, 242, 243, 244, 245 or 246.
The continuous polypeptide of any preceding embodiment, wherein the Trk a ECD polypeptide is glycosylated.
The continuous polypeptide of any one of the preceding embodiments, wherein the Trk a ECD polypeptide comprises at least one glycan moiety.
Embodiment 56. the continuous polypeptide of any one of the preceding embodiments, wherein the Trk a ECD polypeptide is pegylated.
Embodiment 57 the continuous polypeptide of any one of the preceding embodiments, wherein the Trk a ECD polypeptide is pegylated at the glycan, at the primary amine, and/or at the N-terminal alpha-amine.
Embodiment 58. an isolated nucleic acid encoding a contiguous polypeptide according to any of the preceding embodiments.
Embodiment 59. a host cell comprising a nucleic acid according to embodiment 58.
Embodiment 60 a method of producing a polypeptide, the method comprising culturing a host cell according to embodiment 59 and isolating the contiguous polypeptide.
Embodiment 61. a pharmaceutical composition comprising a continuous polypeptide according to any one of embodiments 1 to 57 and a pharmaceutically acceptable carrier.
Embodiment 62 the pharmaceutical composition of embodiment 61, wherein the pharmaceutically acceptable carrier comprises about 5 to about 50mM sodium citrate; about 5 to about 50mM histidine; or about 5 to about 50mM sodium acetate.
Embodiment 63 the pharmaceutical composition of embodiment 61 or embodiment 62, wherein the pharmaceutical composition has a pH of 5 to 6.
Embodiment 64 a method of treating a companion animal species having an NGF-induced disorder comprising administering to the companion animal species a therapeutically effective amount of a continuous polypeptide according to any one of embodiments 1 to 52 or a pharmaceutical composition according to any one of embodiments 61 to 63.
Embodiment 65. a method of treating a companion animal species suffering from pain, comprising administering to the companion animal species a therapeutically effective amount of a continuous polypeptide according to any one of embodiments 1 to 57 or a pharmaceutical composition according to any one of embodiments 61 to 63.
Embodiment 66. the method of embodiment 64 or embodiment 65 wherein the companion animal species is canine, feline or equine.
Embodiment 67. the method of any one of embodiments 64 to 66, wherein the NGF-induced disorder or the pain is chronic pain, acute pain, and/or inflammatory pain.
Embodiment 68. the method of any one of embodiments 64 to 67, wherein the NGF-induced disorder or the pain is osteoarthritis pain, back pain, cancer pain, and/or neuropathic pain.
Embodiment 69 the method of any one of embodiments 64 to 68, wherein the NGF-induced disorder or the pain is pain associated with surgery, a broken bone or fracture, dental treatment, a burn, a cut, and/or childbirth.
Embodiment 70 the method of any one of embodiments 64 to 69, wherein the continuous polypeptide or the pharmaceutical composition is administered parenterally.
Embodiment 71. the method according to any one of embodiments 64 to 70, wherein said continuous polypeptide or said pharmaceutical composition is administered by intramuscular route, intraperitoneal route, intracerobrospinal route, subcutaneous route, intraarterial route, intrasynovial route, intrathecal route or inhalation route.
Embodiment 72 the method of any one of embodiments 64 to 71, wherein the method further comprises administering an NGF kinase inhibitor, a PI3K inhibitor, a ras inhibitor, a CGRP inhibitor, a TNF inhibitor, an IL17 inhibitor, an EGFR inhibitor, and/or a phospholipase C pathway inhibitor.
Embodiment 73. the method according to any one of embodiments 64 to 72, wherein the method further comprises administering one or more pain treatment drugs, such as corticosteroids, non-steroidal anti-inflammatory drugs (NSAIDs), cyclooxygenase inhibitors, opioids and/or cannabinoids.
Embodiment 74 a method of reducing NGF signaling activity in a cell, the method comprising exposing the cell to the continuous polypeptide of any one of embodiments 1 to 57 or the pharmaceutical composition of any one of embodiments 61 to 63 under the following conditions: the conditions allow binding of the contiguous polypeptide to NGF.
Embodiment 75. the method of embodiment 74, wherein the cells are exposed to the continuous polypeptide or the pharmaceutical composition ex vivo.
Embodiment 76. the method of embodiment 74, wherein the cell is exposed to the continuous polypeptide or the pharmaceutical composition in vivo.
Embodiment 77 the method of any one of embodiments 74 to 76, wherein the cell is a canine cell, a feline cell, or an equine cell.
Embodiment 78 a method for detecting NGF in a sample from a companion animal species, the method comprising contacting the sample with the continuous polypeptide according to any one of embodiments 1 to 57 or the pharmaceutical composition according to any one of embodiments 61 to 63 under the following conditions: the conditions allow binding of the contiguous polypeptide to NGF; and detecting whether a complex is formed between the polypeptide and NGF in the sample.
Embodiment 79 the method of embodiment 78, wherein the sample is a biological sample obtained from a canine, a feline, or an equine.
Drawings
FIG.1 shows a sensorgram comparing the affinity of canine and feline TrkA ECD v2 and v3 IgG-Fc fusion proteins for NGF. Irrelevant canine or feline IgG-Fc fusion proteins were used as negative controls.
FIG.2 shows a sensorgram comparing the affinity of the equine TrkA ECD v2 and v3 IgG-Fc fusion proteins for NGF. An irrelevant equine IgG-Fc fusion protein was used as a negative control.
FIG.3 is a graph showing NGF activity neutralization by an exemplary equine TrkA ECD-IgG Fc polypeptide (SEQ ID NO:22) in a TF1 cell proliferation assay performed as described in example 4. Irrelevant monoclonal antibodies were used as negative controls.
FIG.4 is a graph showing NGF concentration in synovium following administration of canine TrkA-Fc polypeptides (1mg/kg and 20mg/kg) in a rat MIA-induced osteoarthritis model.
Figure 5 shows Biacore sensorgrams of binding of canine FcRn to wild-type canine IgG-B Fc polypeptides at various concentrations (12.5, 25, 50, 100, and 200 nM).
Figure 6 shows Biacore sensorgrams of binding of canine FcRn to variant canine IgG-B Fc polypeptide L (23) Y at various concentrations (12.5, 25, 50, 100, and 200 nM).
Figure 7 shows Biacore sensorgrams of binding of canine FcRn to variant canine IgG-B Fc polypeptide L (23) F at various concentrations (12.5, 25, 50, 100, and 200 nM).
Figure 8 shows Biacore sensorgrams of binding of canine FcRn to variant canine IgG-B Fc polypeptide L (23) M at various concentrations (12.5, 25, 50, 100, and 200 nM).
Figure 9 shows Biacore sensorgrams of binding of canine FcRn to variant canine IgG-B Fc polypeptide YTE at various concentrations (12.5, 25, 50, 100 and 200 nM).
FIG.10 is an OctetRed sensorgram for binding of chimeric variant canine IgG-A Fc F00 antibody (A) and IgG-D Fc F00 antibody (B) to canine FcRn compared to chimeric variant canine IgG-A Fc (C) without a Phe mutation and IgG-D Fc (D) without a Phe mutation.
Figure 11 shows the serum pharmacokinetic profile of chimeric variant canine IgG-A F00 antibody ("IgG-A F00"; n ═ 2) and chimeric variant canine IgG-a without a Phe mutation ("IgG-a"; n ═ 2) following subcutaneous administration at 2mg/kg to rats.
Figure 12 is an OctetRed sensorgram for binding of chimeric antibodies with variant canine IgG-B Fc (0Y0, 0YH, 0YY, or 00Y) to canine FcRn compared to chimeric antibodies with wild-type canine IgG-B.
Figure 13 is a graph showing the percentage of antibodies normalized over time resulting from an in vivo pharmacokinetic study in dogs as described in example 19.
DESCRIPTION OF THE SEQUENCES
Table 1 provides a list of certain sequences referred to herein.
Detailed Description
A continuous polypeptide is provided comprising at least one TrkA ECD polypeptide that binds an NGF polypeptide. Methods of producing and purifying the continuous polypeptides are also provided. Methods of treatment using TrkA ECD polypeptides that bind NGF and inhibit NGF-mediated signaling are provided. Such methods include, but are not limited to, methods of treating pain in a companion animal species. Also provided are methods of detecting NGF in a sample from a companion animal species.
For the convenience of the reader, the following definitions of terms used herein are provided.
As used herein, numerical terms such as Kd are calculated based on scientific measurements and are therefore subject to appropriate measurement errors. In some cases, numerical terms may include numbers that are rounded to the nearest significant figure.
As used herein, "a" or "an" means "at least one" or "one or more" unless stated otherwise. As used herein, the term "or" means "and/or," unless otherwise indicated. In the context of several dependent claims, the use of "or" when referring to other claims is only to those claims in the alternative.
Exemplary TrkA Polypeptides
TrkA ECD polypeptides that bind NGF, such as canine, feline, and equine TrkA ECD polypeptides that bind NGF, are provided.
"amino acid sequence" means an amino acid sequence in a protein, and includes amino acid sequences in which one or more amino acid side groups of the sequence have been chemically modified, as well as those amino acid sequences such as: wherein one or more amino acids have been substituted, inserted or deleted relative to the known sequence, but without thereby abrogating the desired properties, such as the ability to bind to the EPO receptor. The amino acid sequence may also be referred to as a peptide, oligopeptide, polypeptide or protein.
As used herein, a "TrkA" or "TrkA polypeptide" is a polypeptide comprising all or a fragment of a tropomyosin receptor kinase a (TrkA) receptor capable of binding NGF.
For example, unless otherwise specified, "TrkA" refers to TrkA polypeptides from any vertebrate source, including mammals such as primates (e.g., humans and cynomolgus monkeys), rodents (e.g., mice and rats), and companion animals (e.g., dogs, cats, and horses). In some embodiments, TrkA is an extracellular domain fragment that binds NGF. In some such embodiments, TrkA may be referred to as TrkA extracellular domain (ECD). In some embodiments, TrkA comprises the amino acid sequence: SEQ ID NO 1, SEQ ID NO 2, SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 5, SEQ ID NO 6, SEQ ID NO 7, SEQ ID NO 8, SEQ ID NO 9, SEQ ID NO 10, SEQ ID NO 11, SEQ ID NO 12, SEQ ID NO 13, SEQ ID NO 14, SEQ ID NO 15, SEQ ID NO 25, SEQ ID NO 26, SEQ ID NO 27, SEQ ID NO 28, SEQ ID NO 29, SEQ ID NO 30, SEQ ID NO 31, SEQ ID NO 32 or SEQ ID NO 33.
As used herein, an "NGF receptor" or "NGFR" is a polypeptide comprising all or part of a low affinity nerve growth factor receptor (also known as tumor necrosis factor receptor superfamily member 16) that binds NGF.
For example, unless otherwise indicated, "NGFR" refers to an NGFR polypeptide from any vertebrate source, including mammals, such as primates (e.g., humans and cynomolgus monkeys), rodents (e.g., mice and rats), and companion animals (e.g., dogs, cats, and horses). In some embodiments, the NGFR is an extracellular domain fragment that binds NGF. In some such embodiments, the NGFR can be referred to as an NGFR extracellular domain (ECD). In some embodiments, the NGFR comprises the amino acid sequence of SEQ ID NO 134, 135, 136, 137, 138 or 139.
The term "companion animal species" refers to an animal suitable for human companion. In some embodiments, the companion animal species is a small mammal such as a canine, feline, dog, cat, horse, rabbit, ferret, guinea pig, rodent, and the like. In some embodiments, the companion animal species is a farm animal such as cattle, swine, and the like.
An "extracellular domain" ("ECD") is the portion of a polypeptide that extends beyond the transmembrane domain into the extracellular space. As used herein, the term "extracellular domain" may comprise an intact extracellular domain or may comprise a truncated extracellular domain lacking one or more amino acids that bind its ligand. The composition of the extracellular domain may depend on the algorithm used to determine which amino acids are in the membrane. For a given protein, different algorithms may predict different extracellular domains, and different systems may express different extracellular domains.
The extracellular domain of the TrkA polypeptide may comprise the entire extracellular domain of TrkA or a truncated extracellular domain that binds NGF. As used herein, the term "extracellular domain of a TrkA polypeptide" or "TrkA ECD" refers to a TrkA polypeptide that does not include a transmembrane domain or a cytoplasmic domain, even if the term follows an open transition word, such as "comprising", "comprises", and the like. In some embodiments, the extracellular domain of the TrkA polypeptide is the extracellular domain of TrkA from a companion species animal. For example, in some embodiments, the extracellular domain of the TrkA polypeptide is derived from a canine TrkA, a feline TrkA, or an equine TrkA. In some embodiments, the extracellular domain of a TRKA polypeptide comprises the amino acid sequence of: SEQ ID NO 2, SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 5, SEQ ID NO 7, SEQ ID NO 8, SEQ ID NO 9, SEQ ID NO 10, SEQ ID NO 12, SEQ ID NO 13, SEQ ID NO 14, SEQ ID NO 15, SEQ ID NO 25, SEQ ID NO 26, SEQ ID NO 27, SEQ ID NO 28, SEQ ID NO 29, SEQ ID NO 30, SEQ ID NO 31, SEQ ID NO 32, SEQ ID NO 33 or any fragment thereof.
The extracellular domain of the NGFR polypeptide can comprise the entire extracellular domain or a truncated extracellular domain of NGF-binding NGF. As used herein, the term "extracellular domain of an NGFR polypeptide" or "NGFR ECD" refers to an NGFR polypeptide that does not comprise a transmembrane or cytoplasmic domain, even if the term follows an open transition word, such as "comprising", "comprises", and the like. In some embodiments, the extracellular domain of the NGFR polypeptide is an extracellular domain of an NGFR polypeptide from an animal of a companion species. For example, in some embodiments, the extracellular domain of an NGFR polypeptide is derived from a canine NGFR, a feline NGFR, or an equine NGFR. In some embodiments, the extracellular domain of the NGFR polypeptide comprises the amino acid sequence of SEQ ID No. 135, SEQ ID No. 137, SEQ ID No. 139, or any fragment thereof.
"wild-type" refers to an unmutated form of a naturally occurring polypeptide or fragment thereof. The wild-type polypeptide may be recombinantly produced.
A "variant" or "analog" is interchangeably referred to herein as a polypeptide that differs from a reference polypeptide by single or multiple amino acid substitutions, deletions, and/or additions, and that substantially retains at least one biological activity of the reference polypeptide.
A "biologically active" entity or an entity having "biological activity" is an entity as follows: it has any function associated with or related to a metabolic or physiological process, and/or has a structural, regulatory, or biochemical function of a naturally occurring molecule. A biologically active polypeptide or fragment thereof includes one that can participate in a biological response including, but not limited to, ligand-receptor interaction or antigen-antibody binding. Biological activity may include improved desired activity or reduced undesired activity. An entity can exhibit biological activity when the entity is involved in a molecular interaction with another molecule, when the entity has therapeutic value to alleviate a disease condition, when the entity has prophylactic value to induce an immune response, and when the entity has diagnostic and/or prognostic value to determine the presence of the molecule.
As used herein, "percent (%) amino acid sequence identity" and "homology" with respect to a polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the particular peptide or polypeptide sequence after aligning the sequences and introducing gaps, if necessary to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for determining percent amino acid sequence identity can be accomplished in a variety of ways well known in the art, for example, using publicly available computer software, such as BLAST, BLAST-2, ALIGN, or MEGALIGNTM(DNASTAR) software. One skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximum alignment over the full length of the sequences being compared.
In some embodiments, a variant has at least about 50% sequence identity to a reference polypeptide after aligning the sequences and introducing gaps, if necessary, to achieve a maximum percent sequence identity and not considering any conservative substitutions as part of the sequence identity. Such variants include, for example, polypeptides in which one or more amino acid residues are added, deleted, or the like, at the N-terminus or C-terminus of the polypeptide. In some embodiments, a variant has at least about 50% sequence identity, at least about 60% sequence identity, at least about 65% sequence identity, at least about 70% sequence identity, at least about 75% sequence identity, at least about 80% sequence identity, at least about 85% sequence identity, at least about 90% sequence identity, at least about 91% sequence identity, at least about 92% sequence identity, at least about 93% sequence identity, at least about 94% sequence identity, at least about 95% sequence identity, at least about 96% sequence identity, at least about 97% sequence identity, at least about 98% sequence identity, or at least 99% sequence identity to the sequence of a reference polypeptide.
As used herein, "a position corresponding to position n" (where n is any number) refers to an amino acid position of the subject polypeptide that aligns with position n of the reference polypeptide after aligning the amino acid sequences of the subject polypeptide and the reference polypeptide and introducing a gap. Alignment for the purpose of determining whether a position of a subject polypeptide corresponds to position n of a reference polypeptide can be achieved in various ways within the skill in the art, e.g., using publicly available computer software such as BLAST, BLAST-2, CLUSTAL OMEGA, ALIGN, or MEGALIGNTM(DNASTAR) software. One skilled in the art can determine appropriate parameters for alignment, including any parameters needed to achieve maximum alignment over the full length of the two sequences being compared. In some embodiments, the subject polypeptide and the reference polypeptide are of different lengths.
A "point mutation" is a mutation involving a single amino acid residue. The mutation may be an amino acid loss, a substitution of one amino acid residue by another amino acid residue, or an insertion of another amino acid residue.
An "amino acid substitution" may include, but is not limited to, the substitution of one amino acid in a polypeptide with another amino acid. Exemplary substitutions are shown in table 2. Amino acid substitutions may be introduced into the molecule of interest and the product screened for a desired activity, e.g., retained/improved receptor binding, reduced immunogenicity, reduced ADCC and/or CDC, or enhanced pharmacokinetics.
Table 2.
Amino acids can be grouped according to common side chain properties:
(1) hydrophobicity: norleucine, Met, Ala, Val, Leu, Ile;
(2) neutral hydrophilicity: cys, Ser, Thr, Asn, Gln;
(3) acidity: asp and Glu;
(4) alkalinity: his, Lys, Arg;
(5) residues that influence chain orientation: gly, Pro;
(6) aromatic: trp, Tyr, Phe.
Non-conservative substitutions will require the exchange of members of one of these classes for another.
As used herein, "amino acid derivative" refers to any amino acid, modified amino acid, and/or amino acid analog, other than one of the 20 common natural amino acids found in humans. Exemplary amino acid derivatives include natural amino acids not found in humans (e.g., selenocysteine and pyrrolysine, which may be found in some microorganisms) and unnatural amino acids. Exemplary amino acid derivatives include, but are not limited to, amino acid derivatives commercially available by chemical product manufacturers (e.g., sigmaldrich. com/chemistry/chemistry-products. html. One or more amino acid derivatives can be incorporated into a polypeptide at a specific position using a translation system that utilizes a host cell, an orthogonal aminoacyl-tRNA synthetase derived from a eubacterial synthetase, an orthogonal tRNA, and an amino acid derivative. For further explanation, see, e.g., U.S. patent No. 9,624,485.
In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution with an amino acid derivative. In some embodiments, the amino acid derivative is an alanine derivative, a cysteine derivative, an aspartic acid derivative, a glutamic acid derivative, a phenylalanine derivative, a glycine derivative, a histidine derivative, an isoleucine derivative, a lysine derivative, a leucine derivative, a methionine derivative, an asparagine derivative, a proline derivative, a glutamine derivative, an arginine derivative, a serine derivative, a threonine derivative, a valine derivative, a tryptophan derivative, or a tyrosine derivative.
In some embodiments, the TrkA ECD polypeptide comprises the amino acid sequence of SEQ ID NO 2, SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 5, SEQ ID NO 7, SEQ ID NO 8, SEQ ID NO 9, SEQ ID NO 10, SEQ ID NO 12, SEQ ID NO 13, SEQ ID NO 14 or SEQ ID NO 15 except for the presence of at least one N-linked glycosylation site that is not present in SEQ ID NO 2, SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 5, SEQ ID NO 7, SEQ ID NO 8, SEQ ID NO 9, SEQ ID NO 10, SEQ ID NO 12, SEQ ID NO 13, SEQ ID NO 14 or SEQ ID NO 15. In some embodiments, the at least one N-linked glycosylation site comprises the sequence asparagine-xaa-serine, wherein xaa is any amino acid except proline. In some embodiments, the at least one N-linked glycosylation site comprises the sequence asparagine-xaa-threonine, wherein xaa is any amino acid except proline. In some embodiments, the at least one N-linked glycosylation site does not overlap with another N-linked glycosylation site.
In some embodiments, the TrkA ECD polypeptide comprises N-linked glycosylation sites at amino acid positions 6-8, 31-33, 84-86, 85-87, 86-88, 88-90, 90-92, 92-94, and/or 94-96 of SEQ ID NO 3, SEQ ID NO 4, SEQ ID NO 8, SEQ ID NO 9, SEQ ID NO 13, or SEQ ID NO 14. In some embodiments, the TrkA ECD polypeptide comprises N-linked glycosylation sites at amino acid positions 4-6, 29-31, 82-84, 83-85, 84-86, 86-88, 89-90, 90-92, and/or 92-94 of SEQ ID NO 5, SEQ ID NO 10, or SEQ ID NO 15.
In some embodiments, the TrkA ECD polypeptide comprises an amino acid other than proline at an amino acid position corresponding to position 30 or position 85 of SEQ ID No. 3, SEQ ID No. 4, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 13, or SEQ ID No. 14. In some embodiments, the TrkA ECD polypeptide comprises an amino acid other than proline at an amino acid position corresponding to position 28 or position 83 of SEQ ID No. 5, SEQ ID No. 10, or SEQ ID No. 15.
In some embodiments, the TrkA ECD polypeptide comprises a valine, glutamic acid, alanine, or isoleucine at an amino acid position corresponding to position 30 and/or position 85 of SEQ ID No. 3, SEQ ID No. 4, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 13, or SEQ ID No. 14. In some embodiments, the TrkA ECD polypeptide comprises a valine, glutamic acid, alanine, or isoleucine at an amino acid position corresponding to position 28 and/or position 83 of SEQ ID NO:5, SEQ ID NO:10, or SEQ ID NO: 15.
In some embodiments, the TrkA ECD polypeptide comprises one or more amino acid modifications listed in table 3, table 4, or table 5, below.
Table 3.
X indicates any amino acid other than proline (e.g. E, V, A, I etc.).
Table 4.
X indicates any amino acid other than proline (e.g. E, V, A, I etc.).
Table 5.
X indicates any amino acid other than proline (e.g. E, V, A, I etc.).
In some embodiments, the TrkA ECD polypeptide comprises one or more additional disulfide bonds. For example, in some embodiments, the TrkA ECD polypeptide comprises a cysteine at a position corresponding to position 7 and position 89 of SEQ ID NO 2,3, 4, 7, 8, 9, 12, 13 or 14. In some embodiments, the TrkA ECD polypeptide comprises a cysteine at a position corresponding to position 5 and position 87 of SEQ ID No. 5, SEQ ID No. 10, or SEQ ID No. 15. In some embodiments, the TrkA ECD polypeptide comprises cysteines at positions 7 and 89 of SEQ ID NO 2,3, 4, 7, 8, 9, 12, 13, or 14. In some embodiments, the TrkA polypeptide comprises cysteines at positions 5 and 87 of SEQ ID NO. 5, SEQ ID NO. 10, or SEQ ID NO. 15.
In some embodiments, the TrkA ECD polypeptide comprises the amino acid sequence of SEQ ID NO 25, SEQ ID NO 26, SEQ ID NO 27, SEQ ID NO 28, SEQ ID NO 29, SEQ ID NO 30, SEQ ID NO 31, SEQ ID NO 32, or SEQ ID NO 33.
As used herein, "glycosylated" refers to a polypeptide having one or more covalently attached glycan moieties.
As used herein, "glycan" or "glycan moiety" refers to a glycoside-linked monosaccharide.
Glycans are attached to glycopeptides in several ways, with N attached to asparagine and O attached to serine and threonine being the most relevant for recombinant therapeutic glycoproteins. N-linked glycosylation occurs at the consensus sequence Asn-Xaa-Ser/Thr, where Xaa can be any amino acid except proline.
As used herein, "sialylation" refers to a polypeptide having one or more covalently attached sialic acid moieties.
Various methods have been developed for producing glycosylated and sialylated proteins. See, e.g., Savinova et al, Applied Biochem & Microbiol.51(8):827-33 (2015).
As used herein, "pegylated" refers to a polypeptide having one or more associated or covalently or non-covalently attached polyethylene glycol (PEG) moieties.
In some embodiments, the TrkA ECD polypeptide is glycosylated. In some embodiments, the TrkA ECD polypeptide comprises at least one glycan moiety attached to an N-linked glycosylation site. In some embodiments, the TrkA ECD polypeptide is sialylated. In some embodiments, the TrkA ECD polypeptide is pegylated. In some embodiments, the TrkA ECD polypeptide is pegylated at the glycan. In some embodiments, the TrkA ECD polypeptide is pegylated at a primary amine. In some embodiments, the TrkA ECD polypeptide is pegylated at the N-terminal alpha amine. In some embodiments, the TrkA ECD polypeptide is glycosylated, sialylated, and/or pegylated.
Exemplary variant IgG Fc polypeptides and fusion molecules
A contiguous polypeptide comprising a TrkA polypeptide may comprise a fusion partner, such as a wild-type or variant IgG Fc polypeptide.
As used herein, "fusion molecule" refers to a molecule comprising one or more "fusion partners". In some embodiments, the fusion partner is covalently linked ("fused"). If both fusion partners are polypeptides, the fusion partner polypeptide can be part of a contiguous amino acid sequence (i.e., a contiguous polypeptide). The first fusion partner polypeptide may be linked to the N-terminus or C-terminus of the second fusion partner. In some embodiments, the fusion partner is translated as a single polypeptide from the coding sequences encoding both fusion partners. The fusion partners may be covalently linked by other means, such as chemical linkage other than peptide bonds. Many known methods of covalently linking a polypeptide to other molecules (e.g., fusion partners) can be used. In other embodiments, the fusion partner is fused via a "linker" consisting of at least one amino acid or chemical moiety.
In some embodiments, the fusion partner is albumin, an albumin binding fragment, or a fragment of an immunoglobulin molecule. The fusion partner may comprise an oligomerization domain, such as the Fc domain of a heavy chain immunoglobulin. In some embodiments, the fusion partner comprises at least one TrkA ECD polypeptide and one IgG Fc polypeptide. In some embodiments, the fusion partner further comprises one or more additional therapeutic polypeptides, such as a NGFR ECD polypeptide. In some embodiments, the TrkA ECD polypeptide may be linked to the N-terminus or C-terminus of the IgG Fc polypeptide.
The term "contiguous polypeptide" is used herein to mean an uninterrupted sequence of amino acids. A contiguous polypeptide is typically translated from a single contiguous DNA sequence. It can be made by genetic engineering, for example, by removing a stop codon from the DNA sequence of a first protein and then appending in frame the DNA sequence of a second protein so that the DNA sequence is expressed as a single protein. Typically, this is accomplished by in-frame cloning of the cDNA into an expression vector along with the existing gene.
"linker" refers to one or more amino acid residues that link a first polypeptide to a second polypeptide.
In some embodiments, the linker is a flexible, non-structural linker. In some embodiments, the linker is a glycine-rich, serine-rich, or glycine and serine-rich linker. In some embodiments, the linker comprises 100%, at least 95%, at least 90%, or at least 85% serine and/or glycine amino acid residues. In some embodiments, the linker is a glycine-rich, serine-rich, or GS-rich flexible nonstructural linker. In some embodiments, the linker comprises amino acids g (gly) and/or s (ser). For example, a linker may comprise G or a repeat of G (e.g., GG, GGG, etc.); GS or a repetitive sequence of GS (e.g., GSGS (SEQ ID NO:143), GSGSGS (SEQ ID NO:144), etc.); GGS or a repetitive sequence of GGS (e.g., GGSGGS (SEQ ID NO:145), GGSGGSGGS (SEQ ID NO:146), etc.); GGGS (SEQ ID NO:147) or a repetitive sequence of GGGS (SEQ ID NO:147) (e.g., GGGSGGGS (SEQ ID NO:148), GGGSGGGSGGGS (SEQ ID NO:149), etc.); GSS or a repetitive sequence of GSS (e.g., GSSGSS (SEQ ID NO:150), GSSGSSGSS (SEQ ID NO:151), etc.); GGSS (SEQ ID NO:152) or repeats of GGSS (SEQ ID NO:152) (e.g., GGSSGGSS (SEQ ID NO:153), GGSSGGSSGGSS (SEQ ID NO:157), etc.); SGGG (SEQ ID NO:158) or a repeat sequence of SGGG (SEQ ID NO:158) (e.g., SGGGSGGGS (SEQ ID NO: 156)).
As used herein, "extension" refers to one or more amino acid residues attached to a polypeptide at the C-terminus or N-terminus.
In some embodiments, the extension portion is flexible. In some embodiments, the extension portion adds flexibility to the polypeptide without interfering with the biological activity of the polypeptide. In some embodiments, the extension portion increases the solubility of the polypeptide. In some embodiments, the extension portion comprises one or more glycine residues. In some embodiments, the extension comprises one glycine residue, two glycine residues, three glycine residues, four glycine residues, five glycine residues, six glycine residues, seven glycine residues, eight glycine residues, or more glycine residues.
As used herein, a "variant IgG Fc" is an IgG Fc polypeptide that differs from a reference IgG Fc polypeptide by single or multiple amino acid substitutions, deletions, and/or additions, and that substantially retains at least one biological activity of the reference IgG Fc polypeptide.
A "fragment crystallizable polypeptide" or "Fc polypeptide" is the portion of an antibody molecule that interacts with effector molecules and cells. It comprises the C-terminal portion of an immunoglobulin heavy chain. As used herein, an Fc polypeptide includes fragments of an Fc domain that have one or more biological activities of an intact Fc polypeptide. In some embodiments, the biological activity of the Fc polypeptide is the ability to bind FcRn. In some embodiments, the biological activity of the Fc polypeptide is the ability to bind C1 q. In some embodiments, the biological activity of the Fc polypeptide is the ability to bind CD 16. In some embodiments, the biological activity of the Fc polypeptide is the ability to bind protein a. An "effector function" of an Fc polypeptide is an action or activity performed in whole or in part by any antibody in response to a stimulus, and may include complement fixation and/or ADCC (antibody-dependent cellular cytotoxicity) induction.
The term "IgX Fc" or "IgX Fc polypeptide" refers to an Fc polypeptide derived from a particular antibody isotype (e.g., IgG, IgA, IgD, IgE, IgM, etc.), wherein "X" represents an antibody isotype. Thus, "IgG Fc" means that the Fc polypeptide originates from a γ chain, "IgA Fc" means that the Fc polypeptide originates from an α chain, "IgD Fc" means that the Fc polypeptide originates from a δ chain, "IgE Fc" means that the Fc polypeptide originates from an epsilon chain, "IgM Fc" means that the Fc polypeptide originates from a μ chain, and the like. In some embodiments, the IgG Fc polypeptide comprises a hinge, CH2, and CH3, but does not comprise CH1 or CL. In some embodiments, the IgG Fc polypeptide comprises CH2 and CH3, but does not comprise CH1, a hinge, or CL. In some embodiments, the IgG Fc polypeptide comprises CH1, a hinge, CH2, CH3, with or without CL. In some embodiments, an IgG Fc polypeptide comprises CH1, a hinge, CH2, and CH3, with or without CL 1. In some embodiments, an Fc polypeptide (e.g., an IgG Fc polypeptide) lacks one or more C-terminal amino acids, such as 1 to 20, 1 to 15, 1 to 10, 1 to 5, or 1 to 2 amino acids, while retaining biological activity. In some embodiments, the biological activity is the ability to bind FcRn. An "effector function" of an Fc polypeptide is an action or activity performed in whole or in part by any antibody in response to a stimulus, and may include complement fixation and/or ADCC (antibody-dependent cellular cytotoxicity) induction. "IgX-NFc" or "IgGXN Fc" means that the Fc polypeptide is derived from a particular subclass of the antibody isotype (e.g., the canine IgG subclass IgG-A, IgG-B, IgG-C or IgG-D; the feline IgG subclass IgG1a, IgG1b or IgG 2; or the equine IgG subclass IgG1, IgG2, IgG3, IgG4, IgG5, IgG6 or IgG7, etc.), wherein "N" means a subclass.
By "hinge" is meant any portion of an Fc polypeptide or variant Fc polypeptide that is proline-rich and comprises at least one cysteine residue located between CH1 and CH2 of the full-length heavy chain constant region.
In some embodiments, the hinge is capable of forming a disulfide bond with a hinge region of a separate Fc polypeptide or with a separate Fc polypeptide within the same hinge region (within the same Fc polypeptide). In some embodiments, the hinge comprises at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten proline residues.
In some embodiments, the IgX or IgXN region is derived from a companion animal such as a dog, cat, or horse. In some embodiments, the IgG region is isolated from a canine gamma heavy chain, such as IgGA, IgGB, IgGC, or IgGD. In some cases, the IgG Fc region is isolated from a feline gamma heavy chain, such as IgG1, IgG2a, or IgG2 b. In other cases, the IgG region is isolated from an equine gamma heavy chain, such as IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, or IgG 7. Polypeptides comprising an Fc region of IgGA, IgGB, IgGC or IgGD can provide higher expression levels in recombinant production systems.
In some embodiments, the IgX Fc polypeptide or IgX-N Fc polypeptide is derived from a companion animal such as a dog, cat, or horse. In some embodiments, the IgG Fc polypeptide is isolated from a canine gamma heavy chain, such as IgG-A, IgG-B, IgG-C or IgG-D. In some cases, the IgG Fc polypeptide is isolated from a feline gamma heavy chain, such as IgG1a, IgG1b, or IgG 2. In other cases, the IgG Fc polypeptide is isolated from an equine gamma heavy chain, such as IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, or IgG 7.
Unless otherwise indicated, the terms "IgX Fc" and "IgX Fc polypeptide" include wild-type IgX Fc polypeptides and variant IgX Fc polypeptides.
In some embodiments, the wild-type IgG Fc polypeptide comprises the amino acid sequence of: SEQ ID NO 34, 35, 36, 37, 38, 39, 70, 71, 72, 73, 74, 75, 76, 77, 86, 87, 88, 89 or 90.
A "variant Fc polypeptide" is an Fc polypeptide that differs from a reference polypeptide by single or multiple unnatural amino acid substitutions, deletions, and/or additions. In some embodiments, the variant Fc-polypeptide retains at least one biological activity of the reference Fc-polypeptide. In some embodiments, a variant Fc polypeptide (e.g., a variant canine IgG-a Fc, a variant canine IgG-C Fc, a variant canine IgG-D Fc, a variant equine IgG2Fc, a variant equine IgG5 Fc, or a variant equine IgG6 Fc) has an activity that is substantially absent from a reference Fc polypeptide. For example, in some embodiments, a variant canine IgG-a Fc, a variant canine IgG-C Fc, a variant canine IgG-D Fc, a variant equine IgG2Fc, a variant equine IgG5 Fc, or a variant equine IgG6Fc binds protein a.
In some embodiments, the variant IgG Fc polypeptide comprises a variant IgG Fc polypeptide of a companion animal species. In some embodiments, the variant IgG Fc polypeptide comprises a variant canine IgG Fc polypeptide, a variant equine IgG Fc polypeptide, or a feline IgG Fc polypeptide.
Exemplary variant IgG Fc polypeptides with modified protein A binding
In some embodiments, the variant IgG Fc polypeptide has a modified protein a binding affinity. In some embodiments, the variant IgG Fc polypeptide has increased binding affinity to protein a. In some embodiments, the variant IgG Fc polypeptide can be purified using protein a column chromatography.
In some embodiments, the variant IgG Fc-polypeptide comprises an amino acid substitution at a position corresponding to position 21, position 23, position 25, position 80, position 205, and/or position 207 of SEQ ID NO: 34. In some embodiments, the variant IgG Fc-polypeptide comprises an amino acid substitution at a position corresponding to position 21, position 23, and/or position 24 of SEQ ID NO: 37. In some embodiments, the variant IgG Fc-polypeptide comprises an amino acid substitution at a position corresponding to position 21, position 23, position 25, position 80, and/or position 207 of SEQ ID NO: 39.
In some embodiments, the variant IgG Fc-polypeptide comprises an amino acid substitution at a position corresponding to position 15 and/or position 203 of SEQ ID NO: 71. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 199 and/or position 200 of SEQ ID NO: 75. In some embodiments, the variant IgG Fc-polypeptide comprises an amino acid substitution at a position corresponding to position 199, position 200, position 201, and/or 202 of SEQ ID NO: 76.
In some embodiments, the variant IgG Fc-polypeptide comprises an amino acid substitution at position 21, position 23, position 25, position 80, position 205, and/or position 207 of SEQ ID NO: 34. In some embodiments, the variant IgG Fc-polypeptide comprises an amino acid substitution at position 21, position 23, and/or position 24 of SEQ ID NO: 37. In some embodiments, the variant IgG Fc-polypeptide comprises an amino acid substitution at position 21, position 23, position 25, position 80, and/or position 207 of SEQ ID NO: 39.
In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at position 15 and/or position 203 of SEQ ID NO: 71. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at position 199 and/or position 200 of SEQ ID NO: 75. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at position 199, position 200, position 201, and/or position 202 of SEQ ID NO: 76.
In some embodiments, the variant IgG Fc-polypeptide comprises a threonine at a position corresponding to position 21 of SEQ ID No. 34, a leucine at a position corresponding to position 23 of SEQ ID No. 34, an alanine at a position corresponding to position 25 of SEQ ID No. 34, a glycine at a position corresponding to position 80 of SEQ ID No. 34, an alanine at a position corresponding to position 205 of SEQ ID No. 34, and/or a histidine at a position corresponding to position 207 of SEQ ID No. 34. In some embodiments, the variant IgG Fc-polypeptide comprises a threonine at a position corresponding to position 21 of SEQ ID No. 37, a leucine at a position corresponding to position 23 of SEQ ID No. 37, and/or an isoleucine at a position corresponding to position 24 of SEQ ID No. 37. In some embodiments, the variant IgG Fc-polypeptide comprises a threonine at a position corresponding to position 21 of SEQ ID No. 39, a leucine at a position corresponding to position 23 of SEQ ID No. 39, an alanine at a position corresponding to position 25 of SEQ ID No. 39, a glycine at a position corresponding to position 80 of SEQ ID No. 39, and/or a histidine at a position corresponding to position 207 of SEQ ID No. 39.
In some embodiments, the variant IgG Fc-polypeptide comprises a threonine or valine at a position corresponding to position 15 of SEQ ID NO:71 and/or a tyrosine or valine at a position corresponding to position 203 of SEQ ID NO: 71. In some embodiments, the variant IgG Fc-polypeptide comprises a leucine at a position corresponding to position 199 of SEQ ID No. 75 and/or a histidine at a position corresponding to position 200 of SEQ ID No. 75. In some embodiments, the variant IgG Fc-polypeptide comprises an isoleucine at a position corresponding to position 199 of SEQ ID No. 76, a histidine at a position corresponding to position 200 of SEQ ID No. 76, an asparagine at a position corresponding to position 201 of SEQ ID No. 76, and/or a histidine at a position corresponding to position 202 of SEQ ID No. 76.
In some embodiments, the variant IgG Fc-polypeptide comprises a threonine at position 21 of SEQ ID No. 34, a leucine at position 23 of SEQ ID No. 34, an alanine at position 25 of SEQ ID No. 34, a glycine at position 80 of SEQ ID No. 34, an alanine at position 205 of SEQ ID No. 34, and/or a histidine at position 207 of SEQ ID No. 34. In some embodiments, the variant IgG Fc-polypeptide comprises a threonine at position 21, a leucine at position 23, and/or an isoleucine at position 24 of SEQ ID NO: 47. In some embodiments, the variant IgG Fc-polypeptide is a threonine at position 21 of SEQ ID No. 39, a leucine at position 23 of SEQ ID No. 39, an alanine at position 25 of SEQ ID No. 39, a glycine at position 80 of SEQ ID No. 39, and/or a histidine at position 207 of SEQ ID No. 39.
In some embodiments, the variant IgG Fc-polypeptide comprises a threonine or valine at position 15 and/or a tyrosine or valine at position 203 of SEQ ID NO 71. In some embodiments, the variant IgG Fc-polypeptide comprises a leucine at position 199 of SEQ ID No. 75 and/or a histidine at position 200 of SEQ ID No. 75. In some embodiments, the variant IgG Fc polypeptide comprises an isoleucine at position 199 of SEQ ID No. 76, a histidine at position 200 of SEQ ID No. 76, an asparagine at position 201 of SEQ ID No. 76, and/or a histidine at position 202 of SEQ ID No. 76.
Exemplary variant IgG Fc polypeptides with modified CD16 binding
In some embodiments, the variant IgG Fc polypeptide has a modified CD16 binding affinity. In some embodiments, the variant IgG Fc polypeptide has reduced binding affinity to CD 16. In some embodiments, the variant IgG Fc can have a reduced ADCC immune response.
In some embodiments, the variant IgG Fc-polypeptide comprises an amino acid substitution at a position corresponding to position 5, position 38, position 39, position 97, and/or position 98 of SEQ ID NO: 35. In some embodiments, the variant IgG Fc-polypeptide comprises an amino acid substitution at a position corresponding to position 5, position 38, position 39, position 97, and/or position 98 of SEQ ID NO: 37.
In some embodiments, the variant IgG Fc-polypeptide comprises an amino acid substitution at position 5, position 38, position 39, position 97, and/or position 98 of SEQ ID NO: 35. In some embodiments, the variant IgG Fc-polypeptide comprises an amino acid substitution at position 5, position 38, position 39, position 97, and/or position 98 of SEQ ID NO: 37.
In some embodiments, the variant IgG Fc-polypeptide comprises a proline at a position corresponding to position 5 of SEQ ID NO:36, a glycine at a position corresponding to position 38 thereof, an arginine at a position corresponding to position 39 thereof, an isoleucine at a position corresponding to position 97 thereof, and/or a glycine at a position corresponding to position 98 thereof. In some embodiments, the variant IgG Fc-polypeptide comprises a proline at a position corresponding to position 5 of SEQ ID NO:37, a glycine at a position corresponding to position 38 thereof, an arginine at a position corresponding to position 39 thereof, an isoleucine at a position corresponding to position 97 thereof, and/or a glycine at a position corresponding to position 98 thereof.
In some embodiments, the variant IgG Fc-polypeptide comprises a proline at position 5, a glycine at position 38, an arginine at position 39, an isoleucine at position 97, and/or a glycine at position 98 of SEQ ID NO: 35. In some embodiments, the variant IgG Fc-polypeptide comprises a proline at position 5, a glycine at position 38, an arginine at position 39, an isoleucine at position 97, and/or a glycine at position 98 of SEQ ID NO: 37.
Exemplary variant IgG Fc polypeptides with modified C1q binding
In some embodiments, the variant IgG Fc polypeptide has a modified C1q binding affinity. In some embodiments, the variant IgG Fc polypeptide has reduced binding affinity to C1 q. In some embodiments, the variant IgG Fc polypeptide can have reduced complement binding. In some embodiments, the variant IgG Fc can have a reduced complement-mediated immune response.
In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 93 of SEQ ID NO: 35. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 93 of SEQ ID NO: 37. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 87 of SEQ ID NO: 70. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 87 of SEQ ID No. 73. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 87 of SEQ ID NO: 74. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 87 of SEQ ID No. 77. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 198 of SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, or SEQ ID NO: 89.
In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at position 93 of SEQ ID NO: 35. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at position 93 of SEQ ID NO: 37. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at position 87 of SEQ ID NO: 70. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at position 87 of SEQ ID NO: 73. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at position 87 of SEQ ID NO: 74. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at position 87 of SEQ ID No. 77. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at position 198 of SEQ ID No. 86, SEQ ID No. 87, SEQ ID No. 88, or SEQ ID No. 89.
In some embodiments, the variant IgG Fc polypeptide comprises an arginine at a position corresponding to position 93 of SEQ ID NO: 35. In some embodiments, the variant IgG Fc polypeptide comprises an arginine at a position corresponding to position 93 of SEQ ID NO: 37. In some embodiments, the variant IgG Fc polypeptide comprises a serine at a position corresponding to position 87 of SEQ ID NO: 70. In some embodiments, the variant IgG Fc polypeptide comprises a serine substitution at a position corresponding to position 87 of SEQ ID No. 73. In some embodiments, the variant IgG Fc polypeptide comprises a serine at a position corresponding to position 87 of SEQ ID No. 74. In some embodiments, the variant IgG Fc polypeptide comprises a serine at a position corresponding to position 87 of SEQ ID No. 77. In some embodiments, the variant IgG Fc polypeptide comprises an alanine at a position corresponding to position 198 of SEQ ID No. 86, SEQ ID No. 87, SEQ ID No. 88, or SEQ ID No. 89.
In some embodiments, the variant IgG Fc polypeptide comprises an arginine at position 93 of SEQ ID NO: 35. In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at position 93 of SEQ ID NO: 37. In some embodiments, the variant IgG Fc polypeptide comprises a serine at position 87 of SEQ ID NO: 70. In some embodiments, the variant IgG Fc polypeptide comprises a serine at position 87 of SEQ ID NO: 73. In some embodiments, the variant IgG Fc polypeptide comprises a serine at position 87 of SEQ ID NO: 74. In some embodiments, the variant IgG Fc polypeptide comprises a serine at position 87 of SEQ ID No. 77. In some embodiments, the variant IgG Fc polypeptide comprises an alanine at position 198 of SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, or SEQ ID NO: 89.
Exemplary variant IgG Fc polypeptides with modified FcRn binding
In some embodiments, the variant IgG Fc polypeptide has a modified neonatal receptor (FcRn) binding affinity. In some embodiments, the variant IgG Fc polypeptide has increased binding affinity for FcRn.
In some embodiments, the variant IgG Fc polypeptide binds to FcRn with greater affinity than a wild-type IgG Fc polypeptide as measured by biofilm layer interference, surface plasmon resonance, or any protein-protein interaction means at a pH in the range of about 5.0 to about 6.5, as measured at a pH of about 5.0, a pH of about 5.2, a pH of about 5.5, a pH of about 6.0, a pH of about 6.2, or a pH of about 6.5.
In some embodiments, the variant IgG Fc polypeptide is less than 5x10-6M, less than 1x10-6M, less than 5x10-7M, less than 1x10-7M, less than 5x10-8M, less than 1x10-8M, less than 5x10-9M, less than 1x10-9M, less than 5x10-10M, less than 1x10- 10M, less than 5x10-11M, less than 1x10-11M, less than 5x10-12M or less than 1x10-12The dissociation constant (Kd) of M binds to FcRn, as measured by biofilm layer interferometry, surface plasmon resonance, or any protein-protein interaction means at a pH in the range of about 5.0 to about 6.5, such as at a pH of about 5.0, a pH of about 5.5, a pH of about 6.0, or a pH of about 6.5.
In some embodiments, the continuous polypeptide comprises a variant IgG Fc polypeptide capable of binding to FcRn with increased affinity relative to a wild-type Fc polypeptide, and wherein the continuous polypeptide has increased serum half-life relative to a continuous polypeptide comprising a wild-type Fc polypeptide.
In some embodiments, the variant IgG Fc polypeptide comprises a tyrosine or phenylalanine at a position corresponding to position 23 of SEQ ID No. 34, SEQ ID No. 35, SEQ ID No. 37, SEQ ID No. 39, SEQ ID No. 70, SEQ ID No. 71, SEQ ID No. 73, SEQ ID No. 74, SEQ ID No. 75, SEQ ID No. 76, SEQ ID No. 77, SEQ ID No. 86, SEQ ID No. 88, or SEQ ID No. 90. In some embodiments, the variant IgG Fc polypeptide comprises a tyrosine at a position corresponding to position 82 of SEQ ID No. 34, SEQ ID No. 35, SEQ ID No. 37, SEQ ID No. 39, SEQ ID No. 70, SEQ ID No. 71, SEQ ID No. 73, SEQ ID No. 74, SEQ ID No. 75, SEQ ID No. 76, SEQ ID No. 77, SEQ ID No. 86, SEQ ID No. 88, or SEQ ID No. 90. In some embodiments, the variant IgG Fc polypeptide comprises a tyrosine at a position corresponding to position 82 of SEQ ID NO 35, SEQ ID NO 37, SEQ ID NO 70, SEQ ID NO 71, SEQ ID NO 73, SEQ ID NO 74, SEQ ID NO 75, SEQ ID NO 76, SEQ ID NO 77, SEQ ID NO 86, SEQ ID NO 88 or SEQ ID NO 90 and a histidine at a position corresponding to position 207 thereof. In some embodiments, the variant IgG Fc polypeptide comprises a tyrosine at a position corresponding to position 82 of SEQ ID NO 35, SEQ ID NO 37, SEQ ID NO 70, SEQ ID NO 71, SEQ ID NO 73, SEQ ID NO 74, SEQ ID NO 75, SEQ ID NO 76, SEQ ID NO 77, SEQ ID NO 86, SEQ ID NO 88 or SEQ ID NO 90 and a tyrosine at a position corresponding to position 207 thereof. In some embodiments, the variant IgG Fc polypeptide comprises a tyrosine at a position corresponding to position 207 of SEQ ID NO 35, SEQ ID NO 37, SEQ ID NO 70, SEQ ID NO 71, SEQ ID NO 73, SEQ ID NO 74, SEQ ID NO 75, SEQ ID NO 76, SEQ ID NO 77, SEQ ID NO 86, SEQ ID NO 88, or SEQ ID NO 90. In some embodiments, the variant IgG Fc-polypeptide comprises a tyrosine at a position corresponding to position 82 of SEQ ID NO:34 or SEQ ID NO:39 and a histidine at a position corresponding to position 208 thereof. In some embodiments, the variant IgG Fc-polypeptide comprises a tyrosine at a position corresponding to position 82 of SEQ ID NO:34 or SEQ ID NO:39 and a tyrosine at a position corresponding to position 208 thereof. In some embodiments, the variant IgG Fc polypeptide comprises a tyrosine at a position corresponding to position 208 of SEQ ID NO:34 or SEQ ID NO: 39.
In some embodiments, the variant IgG Fc polypeptide comprises a tyrosine or phenylalanine at position 23 of SEQ ID NO 34, 35, 37, 39, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90. In some embodiments, the variant IgG Fc polypeptide comprises a tyrosine at position 82 of SEQ ID NO 34, 35, 37, 39, 70, 71, 73, 74, 75, 76, 77, 86, 88 or 90. In some embodiments, the variant IgG Fc polypeptide comprises a tyrosine at position 82 and a histidine at position 207 of SEQ ID No. 35, SEQ ID No. 37, SEQ ID No. 70, SEQ ID No. 71, SEQ ID No. 73, SEQ ID No. 74, SEQ ID No. 75, SEQ ID No. 76, SEQ ID No. 77, SEQ ID No. 86, SEQ ID No. 88, or SEQ ID No. 90. In some embodiments, the variant IgG Fc polypeptide comprises a tyrosine at position 82 and a tyrosine at position 207 of SEQ ID NO 35, SEQ ID NO 37, SEQ ID NO 70, SEQ ID NO 71, SEQ ID NO 73, SEQ ID NO 74, SEQ ID NO 75, SEQ ID NO 76, SEQ ID NO 77, SEQ ID NO 86, SEQ ID NO 88, or SEQ ID NO 90. In some embodiments, the variant IgG Fc polypeptide comprises a tyrosine at position 207 of SEQ ID NO 35, SEQ ID NO 37, SEQ ID NO 70, SEQ ID NO 71, SEQ ID NO 73, SEQ ID NO 74, SEQ ID NO 75, SEQ ID NO 76, SEQ ID NO 77, SEQ ID NO 86, SEQ ID NO 88, or SEQ ID NO 90. In some embodiments, the variant IgG Fc polypeptide comprises a tyrosine at position 82 and a histidine at position 208 of SEQ ID NO:34 or SEQ ID NO: 39. In some embodiments, the variant IgG Fc polypeptide comprises a tyrosine at position 82 and a tyrosine at position 208 of SEQ ID NO:34 or SEQ ID NO: 39. In some embodiments, the variant IgG Fc polypeptide comprises a tyrosine at position 208 of SEQ ID NO:34 or SEQ ID NO: 39.
Exemplary variant IgG Fc polypeptides with modified interchain disulfide bonds
In some embodiments, the variant cat class IgG Fc polypeptide has at least one additional interchain disulfide bond relative to the wild-type cat class IgG Fc polypeptide. In some embodiments, the variant feline IgG Fc polypeptide has at least one additional interchain disulfide bond in the hinge region. In some embodiments, a variant cat class IgG2Fc polypeptide having at least one additional interchain disulfide bond has increased interchain stability relative to a wild-type cat class IgG Fc polypeptide. In some embodiments, the variant IgG polypeptide has at least one amino acid modification to the hinge region relative to a wild-type IgG Fc polypeptide. In some embodiments, the wild-type IgG Fc polypeptide is a wild-type feline or equine IgG Fc polypeptide. In some embodiments, the variant IgG Fc polypeptide comprises a hinge region or a portion of a hinge region from an IgG Fc polypeptide of a different isotype. In some embodiments, the variant IgG Fc polypeptide comprises a hinge region from a wild-type cat class IgG-1a Fc polypeptide, from a wild-type cat class IgG-1b Fc polypeptide, or from a wild-type equine IgG1Fc polypeptide. In some embodiments, the variant IgG2Fc polypeptide has increased recombinant production and/or increased hinge disulfide formation relative to a wild-type IgG Fc polypeptide. In some embodiments, increased recombinant production and/or increased hinge disulfide formation can be determined by SDS-PAGE analysis under reducing and/or non-reducing conditions.
In some embodiments, the variant IgG Fc-polypeptide comprises a cysteine at a position corresponding to position 8, position 9, position 10, position 11, position 12, position 13, position 14, position 15 or position 16 of SEQ ID No. 90. In some embodiments, the variant IgG Fc-polypeptide comprises a cysteine at position 8, position 9, position 10, position 11, position 12, position 13, position 14, position 15, or position 16 of SEQ ID NO: 90.
In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at a position corresponding to position 16 of SEQ ID No. 86, SEQ ID No. 87, SEQ ID No. 88, SEQ ID No. 89, or SEQ ID No. 90. In some embodiments, the variant IgG Fc-polypeptide comprises an amino acid substitution at a position corresponding to position 3 and/or at a position corresponding to position 20 of SEQ ID NO: 72.
In some embodiments, the variant IgG Fc polypeptide comprises an amino acid substitution at position 16 of SEQ ID No. 86, SEQ ID No. 87, SEQ ID No. 88, SEQ ID No. 89, or SEQ ID No. 90. In some embodiments, the variant IgG Fc-polypeptide comprises an amino acid substitution at position 3 and/or at a position corresponding to position 20 of SEQ ID NO: 72.
In some embodiments, the variant IgG Fc polypeptide comprises a proline at a position corresponding to position 16 of SEQ ID No. 86, SEQ ID No. 87, SEQ ID No. 88, SEQ ID No. 89, or SEQ ID No. 90. In some embodiments, the variant IgG Fc-polypeptide comprises a serine at a position corresponding to position 3 and/or a proline at a position corresponding to position 20 of SEQ ID NO: 72.
In some embodiments, the variant IgG Fc polypeptide comprises a proline at position 16 of SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, or SEQ ID NO: 90. In some embodiments, the variant IgG Fc-polypeptide comprises a serine at position 3 and/or a proline at position 20 of SEQ ID No. 72.
In some embodiments, the variant IgG Fc polypeptide comprises the amino acid sequence of: SEQ ID NO 40, SEQ ID NO 41, SEQ ID NO 42, SEQ ID NO 43, SEQ ID NO 44, SEQ ID NO 45, SEQ ID NO 46, SEQ ID NO 47, SEQ ID NO 48, SEQ ID NO 49, SEQ ID NO 50, SEQ ID NO 51, SEQ ID NO 52, SEQ ID NO 53, SEQ ID NO 54, SEQ ID NO 55, SEQ ID NO 56, SEQ ID NO 57, SEQ ID NO 58, SEQ ID NO 59, SEQ ID NO 60, SEQ ID NO 61, SEQ ID NO 62, SEQ ID NO 63, SEQ ID NO 64, SEQ ID NO 65, SEQ ID NO 66, SEQ ID NO 67, SEQ ID NO 68, SEQ ID NO 69, SEQ ID NO 78, SEQ ID NO 79, SEQ ID NO 80, SEQ ID NO 45, SEQ ID NO 47, SEQ ID NO 48, SEQ ID NO 58, SEQ ID NO 59, SEQ ID NO 69, SEQ ID NO 78, SEQ ID NO 79, SEQ ID NO 80, SEQ ID NO, SEQ ID NO 81, SEQ ID NO 82, SEQ ID NO 83, SEQ ID NO 84, SEQ ID NO 85, SEQ ID NO 91, SEQ ID NO 92, SEQ ID NO 93, SEQ ID NO 94, SEQ ID NO 95, SEQ ID NO 96, SEQ ID NO 97, SEQ ID NO 98, SEQ ID NO 99, SEQ ID NO 100, SEQ ID NO 101, SEQ ID NO 102, SEQ ID NO 103, SEQ ID NO 104, SEQ ID NO 105, SEQ ID NO 106, SEQ ID NO 107, SEQ ID NO 108, SEQ ID NO 109, SEQ ID NO 197, SEQ ID NO 198, SEQ ID NO 199, SEQ ID NO 200, SEQ ID NO 201, SEQ ID NO 202, SEQ ID NO 203, SEQ ID NO 204, SEQ ID NO 205, SEQ ID NO, 206, 207, 208, 209 and/or 210.
In some embodiments, a continuous polypeptide comprising at least one TrkA ECD polypeptide (e.g., ECD v1, v2, v3, and/or v4) and a wild-type or variant canine, feline, or equine IgG Fc polypeptide described herein can be prepared based on the following formula:
formula (I): TrkA ECD 1-L1-Fc;
formula (II): Fc-L1-TrkA ECD 1;
formula (III): TrkA ECD 1-L1-Fc-L2-TrkA ECD 2;
formula (IV): TrkA ECD 1-L1-TrkA ECD 2-L2-Fc; or
Formula (V): Fc-L1-TrkA ECD 1-L2-TrkA ECD 2,
wherein TrkA ECD 1 is a first TrkA ECD polypeptide, TrkA ECD 2 is a second TrkA ECD polypeptide (e.g., the same TrkA ECD polypeptide or a different TrkA ECD polypeptide), L1 and L2 are optional linkers; and Fc is a wild-type or variant IgG Fc polypeptide of the companion animal species. Optionally, the contiguous polypeptide comprises a signal sequence. Exemplary constructs of formulae I-V may comprise a third, fourth or fifth, etc. TrkA ECD after or before any of TrkA ECD 1 or TrkA ECD 2. The third, fourth or fifth, etc. TrkA ECD may be the same TrkA ECD polypeptide as TrkA ECD 1 or TrkA ECD 2 or a different TrkA ECD polypeptide.
For example, the contiguous polypeptide may comprise at least one canine TrkA ECD polypeptide (e.g., SEQ ID NO:2, 3, 4, 5, 25, 26, or 27) and wild-type canine IgG polypeptide (e.g., SEQ ID NO:34, 35, 36, 37, 38, or 39), a variant canine IgG-A Fc polypeptide (e.g., SEQ ID NO:40, 43, 199, or 200), a variant canine IgG-B Fc polypeptide (e.g., SEQ ID NO:46, 48, 49, 50, 51, 52, 53, 54, 55, 64, 65, 66, 67, 197, 198, 203, 204, 205, 206, 207, 208, 209, or 210), a variant canine IgG-C Fc polypeptide (e.g., SEQ ID NO:41, 44, 47, 56, 57, 58, 59, 60, 61, 52, 63, 68, or 69), or a variant IgG-D Fc polypeptide (e.g., SEQ ID NO:42, 45, 25, 26, or 27), 201 or 202) as described herein.
The contiguous polypeptide can comprise at least one cat TrkA ECD polypeptide (e.g., SEQ ID NOs: 7, 8, 9, 10, 28, 29, or 30) and a wild-type cat IgG Fc polypeptide (e.g., 86, 87, 88, 89, or 90), a variant cat IgG1a Fc polypeptide (e.g., SEQ ID NOs: 91, 92, 96, or 97), a variant cat IgG1b Fc polypeptide (e.g., SEQ ID NOs: 93, 94, 98, or 99), or a variant cat IgG2Fc polypeptide (e.g., SEQ ID NOs: 95, 100, or 107), as described herein.
The contiguous polypeptide may comprise at least one equine TrkA ECD polypeptide (e.g., SEQ ID NO:12, 13, 14, 15, 31, 32, or 33) and a wild-type equine IgG Fc polypeptide (e.g., SEQ ID NO:70, 71, 72, 73, 74, 75, 76, or 77), a variant equine IgG1Fc polypeptide (e.g., SEQ ID NO:82), a variant equine IgG2Fc polypeptide (e.g., SEQ ID NO:78, 79, 101, 102, 103, 104, 105, 106, 108, or 109), a variant equine IgG3 Fc polypeptide (e.g., SEQ ID NO:83), a variant equine IgG4 Fc polypeptide (e.g., SEQ ID NO:84), a variant equine IgG5 Fc polypeptide (e.g., SEQ ID NO:80), a variant equine IgG6Fc polypeptide (e.g., SEQ ID NO:81), or a variant equine IgG7Fc polypeptide (e.g., SEQ ID NO: 85).
In some embodiments, the contiguous polypeptide comprising a TrkA ECD polypeptide may further comprise at least one NGFR ECD polypeptide. In some embodiments, the NGFR ECD polypeptide comprises the amino acid sequence of SEQ ID NO:135, SEQ ID NO:137, and/or SEQ ID NO: 139.
In some embodiments, the contiguous polypeptide comprises the amino acid sequence of: 16, 17, 18, 19, 20, 21, 22, 23, 24, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 130, 131, 132, 111, 140, 141, 142, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 186, 187, 188, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 236, 233, 234, 235, 236, 238, 239, 240, 241, 242, 243, 244, 245, 246.
The molecules can be further constructed using the following formats: r- (linker) -Fc-TrkA. R may be any protein, such as TNFR, IL13R, IL4R, IL17R, and the like. The bispecific format may provide additional therapeutic benefits.
TrkA may also be fused to an antibody, for example to the C-terminus of the IgG heavy chain. The antibody may be anti-TNF, anti-CGRP, anti-IL 17, anti-IL 4R, anti-EGFR, etc. The antibody fusions may have an enhanced effect on therapy.
Exemplary TrkA ECD polypeptide expression and production
Polynucleotide sequences encoding all or part (e.g., the extracellular domain) of a TrkA polypeptide with or without a signal sequence are provided. If a homologous signal sequence (i.e., the signal sequence of TRKA) is not used in the construction of the nucleic acid molecule, another signal sequence may be used, for example, any of the signal sequences described in PCT US 06/02951.
Typically, the nucleotide sequence encoding the polypeptide of interest (e.g., a TrkA polypeptide) is inserted into an expression vector suitable for expression in the host cell of choice.
A "vector" is a plasmid that can be used to transfer a DNA sequence from one organism to another or to express a gene of interest. Vectors typically include an origin of replication and regulatory sequences that regulate the expression of the gene of interest, and may or may not carry a selectable marker gene, such as an antibiotic resistance gene. The vector is suitable for the host cell in which it is to be expressed. When the gene of interest is present in a vector, the vector may be referred to as a "recombinant vector".
"host cell" refers to a cell that may be or has been the recipient of a vector or isolated polynucleotide. The host cell may be a prokaryotic cell or a eukaryotic cell. Exemplary embodiments of the inventionEukaryotic cells include mammalian cells, such as primate or non-primate cells; fungal cells, such as yeast; a plant cell; and insect cells. Non-limiting exemplary mammalian cells include, but are not limited to, NS0 cells, PER.Cells (Crucell), 293 cells and CHO cells, and derivatives thereof, such as 293-6E, DG44, CHO-S and CHO-K cells. Host cells include progeny of a single host cell, and the progeny may not necessarily be identical (in morphology or in genomic DNA complement) to the original parent cell due to natural, accidental, or deliberate mutation. Host cells include cells transfected in vivo with one or more polynucleotides encoding one or more amino acid sequences provided herein.
The term "isolated" as used herein refers to a molecule that has been separated from at least some components that are typically found or produced in nature. For example, a polypeptide is said to be "isolated" when it is separated from at least some of the components of the cell from which it was produced. When a polypeptide is secreted by a cell following expression, the supernatant containing the polypeptide is physically separated from the cell in which it was produced, and is considered to be "isolating" the polypeptide. Similarly, a polynucleotide is said to be "isolated" when it is not part of a larger polynucleotide (such as genomic or mitochondrial DNA in the case of a DNA polynucleotide) that is typically found in nature, or is separated from at least some of the components of the cell that produces it, such as in the case of an RNA polynucleotide. Thus, a DNA polynucleotide contained in a vector within a host cell may be referred to as "isolated".
In some embodiments, the TrkA polypeptide is separated using chromatography (e.g., size exclusion chromatography, ion exchange chromatography, protein a column chromatography, hydrophobic interaction chromatography, and CHT chromatography).
The terms "label" and "detectable label" mean a moiety attached to the TrkA polypeptide to render it detectable. In some embodiments, the label is a detectable label that can produce a signal that can be detected by visual or instrumental means, e.g.Incorporation of radiolabeled amino acids or attachment of biotin moieties to polypeptides, which can be detected by labeled avidin (e.g., streptavidin containing a fluorescent label or enzymatic activity that can be detected by optical or colorimetric methods). Examples of labels for polypeptides include, but are not limited to, the following: a radioisotope or radionuclide (e.g.,3H、14C、35S、90Y、99Tc、111In、125I、131I、177Lu、166ho or153Sm); chromogens, fluorescent labels (e.g., FITC, rhodamine, lanthanide phosphors), enzyme labels (e.g., horseradish peroxidase, luciferase, alkaline phosphatase); a chemiluminescent label; a biotin group; a predetermined polypeptide epitope (e.g., a leucine zipper pair sequence, a binding site for a second antibody, a metal binding domain, an epitope tag) recognized by the second reporter molecule; and magnetic agents such as gadolinium chelates. Representative examples of labels commonly used in immunoassays include light-generating moieties such as acridinium compounds; and a moiety that produces fluorescence, such as fluorescein. In this regard, the moiety may not be detectably labeled by itself, but may become detectable upon reaction with yet another moiety.
Exemplary TrkA polypeptides as decoy receptor traps
The TRKA polypeptides of the invention may act as decoy receptors for capturing NGF and inhibiting its interaction with NGF and TRKA on the cell surface. Decoy receptors (such as those of the present invention) recognize their ligands with high affinity and specificity, but are structurally incapable of signaling. They compete with the wild-type receptor for ligand binding and participate in ligand/receptor interactions, thereby modulating the activity or number of functional receptors and/or cellular activity downstream of the receptor. Decoy receptors can act as molecular traps for agonist ligands and thereby inhibit ligand-induced receptor activation.
As used herein, "NGF" refers to any native NGF that results from the expression and processing of NGF in a cell. Unless otherwise indicated, the term includes NGF from any vertebrate source, including mammals such as primates (e.g., humans and cynomolgus monkeys) and rodents (e.g., mice and rats) and companion animals (e.g., dogs, cats, and horses). The term also includes naturally occurring NGF variants, such as splice variants or allelic variants.
The invention provides TrkA ECD polypeptides as therapeutic agents. The TrkA ECD polypeptides of the invention bind NGF (which has been shown to be associated with chronic pain or inflammatory pain) as described in more detail herein. In various embodiments, the TrkA polypeptide may bind NGF with high affinity. In various embodiments, the TrkA polypeptide may interfere with NGF signaling.
The term "affinity" means the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., a receptor) and its binding partner (e.g., a ligand). The affinity of a molecule X for its partner Y can generally be determined by the dissociation constant (K)D) And (4) showing. Affinity can be measured by common methods known in the art, such as immunoblotting, ELISA KD, KinEx a, Biofilm Layer Interferometry (BLI), or surface plasmon resonance devices.
The term "KD”、“Kd"," Kd "or" Kd value "are used interchangeably to refer to the equilibrium dissociation constant for the receptorbody-ligand interaction. In some embodiments, according to the supplier's instructions, use such asBiosensors such as the system (Pall ForteBio LLC, friemont, ca) measure K of fusion molecules and their ligands by using biofilm layer interferometryd. Briefly, biotinylated antigen was bound to the sensor tip and association of the fusion molecules was monitored for 90 seconds and dissociation for 600 seconds. The buffer used for the dilution and binding steps was 20mM phosphate, 150mM NaCl, pH 7.2. The buffer only blank curve was subtracted to correct for any drift. Data were fitted to a 2:1 binding model using ForteBio data analysis software to determine the association rate constant (k)on) Dissociation rate constant (k)off) And Kd. Equilibrium dissociation constant (K)d) Is calculated as koff/konThe ratio of (a) to (b). The term "kon" refers to the rate constant for the association of molecule X with its partner Y, while the term "koff" refers to the rate constant for dissociation of the molecule X or partner Y from the molecule X/partner Y complex.
The term "binding" to a substance is a term well known in the art, and methods of determining such binding are also well known in the art. A molecule is said to exhibit "binding" if it reacts with, associates with, or has affinity for a particular cell or substance, and the reaction, association, or affinity can be detected by one or more methods known in the art (e.g., immunoblotting, ELISA KD, KinEx a, Biofilm Layer Interferometry (BLI), surface plasmon resonance device, etc.).
"surface plasmon resonance" refers to an optical phenomenon that allows analysis of real-time biospecific interactions by detecting changes in protein concentration within a biosensor matrix, for example using BIAcoreTMSystems (BIAcore International AB, a GE Healthcare, Uppsala, Sweden and Piscataway, N.J.). For further explanation, see Jonsson et al (1993) Ann.biol.Clin.51: 19-26.
"biofilm layer interferometry" refers to an optical analysis technique that analyzes the interference pattern of light reflected from the immobilized protein layer on the biosensor tip and internal reference layer. The variation in the number of molecules bound to the biosensor tip causes a shift in the interference pattern that can be measured in real time. A non-limiting exemplary device for biofilm interferometry isSystem (Pall ForteBio LLC). See, e.g., Abdiche et al, 2008, anal. biochem.377: 209-glass 277.
In some embodiments, the TrkA polypeptide is less than 5x10 as measured by biofilm layer interferometry-6M, less than 1x10-6M, less than 5x10-7M, less than 1x10-7M, less than 5x10-8M, smallAt 1x10-8M, less than 5x10-9M, less than 1x10-9M, less than 5x10-10M, less than 1x10-10M, less than 5x10-11M, less than 1x10-11M, less than 5x10-12M or less than 1x10-12The dissociation constant (Kd) of M binds to canine NGF, feline NGF, equine NGF or human NGF. In some embodiments, the TRKA polypeptide binds canine NGF, feline NGF, or equine NGF with a Kd as measured by biofilm layer interferometry as follows: at 5x10-6M and 1x10-6Between M, at 5x10-6M and 5x10-7Between M, at 5x10-6M and 1x10-7Between M, at 5x10-6M and 5x10-8M、5x10-6M and 1x10-8Between M, at 5x10-6M and 5x10-9Between M, at 5x10-6M and 1x10-9Between M, at 5x10-6M and 5x10-10Between M, at 5x10-6M and 1x10-10Between M, at 5x10-6M and 5x10-11Between M, at 5x10-6M and 1x10-11Between M, at 5x10-6M and 5x10-12Between M, at 5x10-6M and 1x10-12Between M, at 1x10-6M and 5x10-7Between M, at 1x10-6M and 1x10-7Between M, at 1x10-6M and 5x10-8M、1x10-6M and 1x10-8Between M, at 1x10-6M and 5x10-9Between M, at 1x10-6M and 1x10-9Between M, at 1x10-6M and 5x10-10Between M, at 1x10-6M and 1x10-10Between M, at 1x10-6M and 5x10-11Between M, at 1x10-6M and 1x10-11Between M, at 1x10-6M and 5x10-12Between M, at 1x10-6M and 1x10-12Between M, at 5x10-7M and 1x10-7Between M, at 5x10-7M and 5x10-8M、5x10-7M and 1x10-8Between M, at 5x10-7M and 5x10-9Between M, at 5x10-7M and 1x10-9Between M, at 5x10-7M and 5x10-10Between M, at 5x10-7M and 1x10-10Between M, at 5x10-7M and 5x10- 11Between M, at 5x10-7M and 1x10-11Between M, at 5x10-7M and 5x10-12Between M, at 5x10-7M and 1x10-12Between M, at 1x10-7M and 5x10-8M,1x10-7M and 1x10-8Between M, at 1x10-7M and 5x10-9Between M, at 1x10-7M and 1x10-9Between M, at 1x10-7M and 5x10-10Between M, at 1x10-7M and 1x10-10Between M, at 1x10-7M and 5x10-11Between M, at 1x10- 7M and 1x10-11Between M, at 1x10-7M and 5x10-12Between M, at 1x10-7M and 1x10-12Between M, at 5x10-8M and 1x10-8Between M, at 5x10-8M and 5x10-9Between M, at 5x10-8M and 1x10-9Between M, at 5x10-8M and 5x10-10Between M, at 5x10-8M and 1x10-10Between M, at 5x10-8M and 5x10-11Between M, at 5x10-8M and 1x10-11Between M, at 5x10-8M and 5x10- 12Between M, at 5x10-8M and 1x10-12M、1x10-8M and 5x10-9Between M, at 1x10-8M and 1x10-9Between M, at 1x10-8M and 5x10-10Between M, at 1x10-8M and 1x10-10Between M, at 1x10-8M and 5x10-11Between M, at 1x10-8M and 1x10-11Between M, at 1x10-8M and 5x10-12Between M, at 1x10-8M and 1x10-12Between M, at 5x10-9M and 1x10-9Between M, at 5x10- 9M and 5x10-10Between M, at 5x10-9M and 1x10-10Between M, at 5x10-9M and 5x10-11Between M, at 5x10-9M and 1x10- 11Between M, at 5x10-9M and 5x10-12Between M, at 5x10-9M and 1x10-12Between M, at 1x10-9M and 5x10-10Between M, at 1x10-9M and 1x10-10Between M, at 1x10-9M and 5x10-11Between M, at 1x10-9M and 1x10-11Between M, at 1x10-9M and 5x10-12Between M, at 1x10-9M and 1x10-12Between M, at 5x10-10M and 1x10-10Between M, at 5x10-10M and 5x10-11Between M, at 1x10-10M and 5x10-11M、1x10-10M and 1x10-11Between M, at 1x10-10M and 5x10-12Between M, at 1x10-10M and 1x10-12Between M, at 5x10-11M and 1x10-12Between M, at 5x10-11M and 5x10-12Between M, at 5x10-11M and 1x10-12Between M, at 1x10-11M and 5x10-12M is at 1x10-11M and 1x10-12M is greater than or equal to the total weight of the composition. In some embodiments, the TRKA polypeptide binds canine NGF, feline NGF, and/or equine NGF.
By "reduce" or "inhibit" is meant a decrease, or arrest in activity, function, or amount as compared to a reference. In some embodiments, "reduce" or "inhibit" means the ability to cause an overall reduction of 20% or more. In some embodiments, "reduce" or "inhibit" means the ability to cause an overall reduction of 50% or more. In some embodiments, "reduce" or "inhibit" means the ability to cause an overall reduction of 75%, 85%, 90%, 95%, or more. In some embodiments, the amount is inhibited or reduced over a period of time relative to a control dose (e.g., placebo) over the same period of time. As used herein, "reference" refers to any sample, standard, or level used for comparison purposes. The reference can be obtained from healthy or non-diseased samples. In some instances, the reference is obtained from an undiseased or untreated sample of the companion animal. In some examples, the reference is obtained from one or more healthy animals of a particular species, but not the animal being tested or treated.
As used herein, the term "significant decrease" means a sufficiently high degree of decrease between a numerical value and a reference numerical value such that one of skill in the art would consider the difference between the two values to be statistically significant in the context of the biological characteristic measured by the value. In some embodiments, the significant decrease is a numerical decrease of greater than any of about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, or 100% compared to the reference value.
In some embodiments, the TrkA polypeptide may reduce NGF signaling in an companion animal species by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% as compared to NGF signaling in the absence of the fusion molecule. In some embodiments, signaling is measured by a decrease in NGF-dependent TF-1 cell proliferation. In some embodiments, the reduction in NGF signaling or reduction in proliferation is between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 10% and 60%, between 10% and 70%, between 10% and 80%, between 10% and 90%, between 10% and 100%, between 15% and 20%, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 15% and 60%, between 15% and 70%, between 15% and 80%, between 15% and 90%, between 15% and 100%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 20% and 45%, between 15% and 90%, between 15% and 100%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 20% and 45%, or between 45% and 45% Between 20% and 50%, between 20% and 60%, between 20% and 70%, between 20% and 80%, between 20% and 90%, between 20% and 100%, between 25% and 30%, between 25% and 35%, between 25% and 40%, between 25% and 45%, between 25% and 50%, between 25% and 60%, between 25% and 70%, between 25% and 80%, between 25% and 90%, between 25% and 100%, between 30% and 35%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 30% and 60%, between 30% and 70%, between 30% and 80%, between 30% and 90%, between 30% and 100%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 35% and 60%, between 35% and 70%, between 35% and 80%, between 35% and 90%, between 35% and 100%, between 35% and 45%, between 35% and 50%, between 35% and 60%, between 35% and 70%, between 35% and 80%, between 35% and 90%, between 35% and 100% Between 40% and 45%, between 40% and 50%, between 40% and 60%, between 40% and 70%, between 40% and 80%, between 40% and 90%, between 40% and 100%, between 45% and 50%, between 45% and 60%, between 45% and 70%, between 45% and 80%, between 45% and 90%, between 45% and 100%, between 50% and 60%, between 50% and 70%, between 50% and 80%, between 50% and 90%, between 50% and 100%, between 60% and 70%, between 60% and 80%, between 60% and 90%, between 60% and 100%, between 70% and 80%, between 70% and 90%, between 70% and 100%, between 80% and 90%, or between 90% and 100%.
Exemplary pharmaceutical compositions
The terms "pharmaceutical formulation" and "pharmaceutical composition" refer to a formulation in a form effective for the biological activity of one or more active ingredients and free of other components having unacceptable toxicity to a subject to which the formulation is administered.
By "pharmaceutically acceptable carrier" is meant a non-toxic solid, semi-solid, or liquid filler, diluent, encapsulating material, formulation aid or carrier conventional in the art for use with a therapeutic agent that together constitute a "pharmaceutical composition" for administration to a subject. Pharmaceutically acceptable carriers are non-toxic to recipients at the dosages and concentrations employed, and are compatible with other ingredients of the formulation. Pharmaceutically acceptable carriers are suitable for the formulation used. Examples of pharmaceutically acceptable carriers include alumina; aluminum stearate; lecithin; serum proteins such as human serum albumin, canine or other animal albumin, and the like; buffers such as phosphate, citrate, tromethamine or HEPES buffers; glycine; sorbic acid; potassium sorbate; partial glyceride mixtures of saturated vegetable fatty acids; water; salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, or magnesium trisilicate; polyvinylpyrrolidone, a cellulose-based substance; polyethylene glycol; sucrose; mannitol; or amino acids including, but not limited to, arginine.
The pharmaceutical composition may be stored in lyophilized form. Thus, in some embodiments, the method of preparation comprises a lyophilization step. The lyophilized composition can then be reconstituted prior to administration to a dog, cat, or horse, typically as an aqueous composition suitable for parenteral administration. In other embodiments, in particular when the fusion molecule is highly stable to thermal and oxidative denaturation, the pharmaceutical composition may be stored as a liquid, i.e. as an aqueous composition, which may be administered directly or with appropriate dilution to a dog, cat or horse. The lyophilized composition can be reconstituted with sterile water for injection (WFI). Bacteriostatic agents such as benzyl alcohol may be included. Accordingly, the present invention provides pharmaceutical compositions in solid or liquid form.
When administered, the pH of the pharmaceutical composition may be in the range of about pH 5 to about pH 8. If the compositions of the present invention are used for therapeutic purposes, they are sterile. Sterility can be achieved by any of several methods known in the art, including filtration through sterile filtration membranes (e.g., 0.2 micron membranes). The sterility may be maintained with or without the use of antibacterial agents.
Exemplary uses of TrkA Polypeptides and pharmaceutical compositions
TrkA polypeptides or pharmaceutical compositions comprising TrkA polypeptides of the present invention may be used to treat NGF-induced disorders. As used herein, "NGF-induced disorder" means a disease associated with, caused by, or characterized by an elevated level or altered distribution of NGF. Such NGF-induced disorders include, but are not limited to, osteoarthritis pain, cancer pain, lower back pain. In some embodiments, the NGF-induced disorder is chronic pain or inflammatory pain. NGF-induced disorders can be exhibited in companion animals including, but not limited to, canines, felines, or equines.
As used herein, "treatment" is a means for obtaining a beneficial or desired clinical result. As used herein, "treatment" includes any administration or use of a disease therapeutic agent in a mammal (including companion animals). For purposes of this disclosure, beneficial or desired clinical results include, but are not limited to, any one or more of the following: alleviating one or more symptoms, reducing the extent of disease, preventing or delaying spread of disease, preventing or delaying relapse of disease, delaying or slowing progression of disease, ameliorating the disease state, inhibiting disease or disease progression, inhibiting or slowing disease or disease progression, arresting disease progression, and alleviating (whether partial or total). "treating" also encompasses reducing the pathological consequences of a proliferative disease. The methods provided herein contemplate any one or more of these therapeutic aspects. Consistent with the above, the term treatment does not require one hundred percent removal of all aspects of the disorder.
In some embodiments, a TrkA polypeptide or a pharmaceutical composition comprising the same may be used to treat an NGF-induced disorder according to the methods herein. In some embodiments, a TrkA polypeptide or pharmaceutical composition is administered to a companion animal, such as a canine, feline, or equine, to treat an NGF-induced disorder.
The "therapeutically effective amount" of a substance/molecule, agonist or antagonist can vary depending on factors such as the type of disease being treated, the disease state, the severity and course of the disease, the type of treatment objective, any previous treatment, clinical history, response to previous treatment, the judgment of the attending veterinarian, the age, sex and weight of the animal, and the ability of the substance/molecule, agonist or antagonist to elicit a desired response in the animal. A therapeutically effective amount is also an amount that has a therapeutically beneficial effect over any toxic or detrimental effects of the substance/molecule, agonist or antagonist. A therapeutically effective amount may be delivered in one or more administrations. A "therapeutically effective amount" is an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result.
In some embodiments, the TrkA polypeptide or the pharmaceutical composition comprising the TrkA polypeptide is administered parenterally, by subcutaneous administration, intravenous infusion, or intramuscular injection. In some embodiments, the TrkA polypeptide or the pharmaceutical composition comprising the TrkA polypeptide is administered as a bolus or by continuous infusion over a period of time. In some embodiments, the TrkA polypeptide or the pharmaceutical composition comprising the TrkA polypeptide is administered by intramuscular, intraperitoneal, intracerobrospinal, subcutaneous, intraarterial, intrasynovial, intrathecal, or inhalation routes.
The TrkA polypeptide described herein may be administered in an amount ranging from 0.1mg/kg body weight to 100mg/kg body weight per dose. In some embodiments, the TrkA fusion may be administered in an amount ranging from 0.5mg/kg body weight to 50mg/kg body weight per dose. In some embodiments, the TrkA fusion may be administered in an amount ranging from 1mg/kg body weight to 10mg/kg body weight per dose. In some embodiments, the fusion molecule can be administered in an amount within the range of 0.5mg/kg body weight to 100mg/kg body weight, within the range of 1mg/kg body weight to 100mg/kg body weight, within the range of 5mg/kg body weight to 100mg/kg body weight, within the range of 10mg/kg body weight to 100mg/kg body weight, within the range of 20mg/kg body weight to 100mg/kg body weight, within the range of 50mg/kg body weight to 100mg/kg body weight, within the range of 1mg/kg body weight to 10mg/kg body weight, within the range of 5mg/kg body weight to 10mg/kg body weight, within the range of 0.5mg/kg body weight to 10mg/kg body weight, or within the range of 5mg/kg body weight to 50mg/kg body weight.
The TrkA polypeptide or pharmaceutical composition comprising the TrkA polypeptide may be administered to the companion animal at one time or over a series of treatments. For example, a TrkA polypeptide or a pharmaceutical composition comprising TrkA may be administered at least once, more than once, at least twice, at least three times, at least four times, or at least five times.
In some embodiments, the dose is administered once a week for at least two or three consecutive weeks, and in some embodiments, the treatment cycle is repeated two or more times, optionally interspersed with one or more weeks of no treatment. In other embodiments, the therapeutically effective dose is administered once daily for two to five consecutive days, and in some embodiments, the treatment cycle is repeated two or more times, optionally interspersed with one or more days of non-treatment.
Administration "in combination with" one or more other therapeutic agents includes simultaneous (concurrent) and sequential or sequential administration in any order. The term "concurrently" is used herein to refer to the administration of two or more therapeutic agents, wherein at least a portion of the administrations overlap in time or the administration of one therapeutic agent occurs within a very short time relative to the administration of the other therapeutic agent. For example, the two or more therapeutic agents are administered at a time interval of no more than about a specified number of minutes. The term "sequentially" is used herein to refer to the administration of two or more therapeutic agents, wherein the administration of one or more agents is discontinued followed by the continued administration of one or more other agents, or wherein the administration of one or more other agents precedes the administration of one or more agents. For example, the two or more therapeutic agents are administered at intervals of time greater than about the specified number of minutes. As used herein, the term "in conjunction with" refers to the administration of one mode of treatment in addition to another mode of treatment. Thus, "in conjunction with … …" refers to administration of one mode of treatment to an animal before, during, or after administration of another mode of treatment to the animal.
In some embodiments, the methods comprise administering an NGF kinase inhibitor, a PI3K inhibitor, a ras inhibitor, and/or a phospholipase C pathway inhibitor in combination with a TrkA polypeptide or a pharmaceutical composition comprising a TrkA polypeptide. In some embodiments, the method further comprises administering one or more pain treatment drugs, such as corticosteroids, non-steroidal anti-inflammatory drugs (NSAIDs), cyclooxygenase inhibitors, opioids and/or cannabinoids.
Provided herein are methods of exposing a cell with a TrkA polypeptide or a pharmaceutical composition comprising a TrkA polypeptide under conditions that permit binding to NGF. In some embodiments, the cells are exposed ex vivo to a TrkA polypeptide or pharmaceutical composition. In some embodiments, the cell is exposed to a TrkA polypeptide or pharmaceutical composition in vivo. In some embodiments, the cell is exposed to a TrkA polypeptide. In some embodiments, the cells are exposed to the TrkA polypeptide or the pharmaceutical composition under the following conditions: the conditions allow the fusion molecule to bind extracellular NGF. In some embodiments, the cells may be exposed to the TrkA polypeptide or the pharmaceutical composition in vivo by any one or more of the methods of administration described herein, including, but not limited to, intraperitoneal, intramuscular, intravenous injection into the body of the subject. In some embodiments, cells may be exposed ex vivo to the TrkA polypeptide or the pharmaceutical composition by: exposing the cell to a culture medium comprising the fusion molecule or the pharmaceutical composition. In some embodiments, the permeability of the cell membrane can be affected using any number of methods understood by those skilled in the art (e.g., electroporating the cells or exposing the cells to a solution containing calcium chloride) prior to exposing the cells to a medium comprising the fusion molecule or the pharmaceutical composition.
In some embodiments, the exposure results in a decrease in NGF signaling function of the cell. In some embodiments, the TrkA polypeptide may reduce NGF signaling in a cell by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% as compared to NGF signaling function in the absence of the TrkA polypeptide. In some embodiments, the reduction in NGF signaling or reduction in TF-1 proliferation is between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 10% and 60%, between 10% and 70%, between 10% and 80%, between 10% and 90%, between 10% and 100%, between 15% and 20%, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 15% and 60%, between 15% and 70%, between 15% and 80%, between 15% and 90%, between 15% and 100%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 15% and 80%, between 15% and 90%, between 15% and 100%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, or more, Between 20% and 45%, between 20% and 50%, between 20% and 60%, between 20% and 70%, between 20% and 80%, between 20% and 90%, between 20% and 100%, between 25% and 30%, between 25% and 35%, between 25% and 40%, between 25% and 45%, between 25% and 50%, between 25% and 60%, between 25% and 70%, between 25% and 80%, between 25% and 90%, between 25% and 100%, between 30% and 35%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 30% and 60%, between 30% and 70%, between 30% and 80%, between 30% and 90%, between 30% and 100%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 35% and 60%, between 35% and 70%, between 35% and 80%, between 35% and 90%, or between 35% and 40%, or a combination of the above, Between 35% and 100%, between 40% and 45%, between 40% and 50%, between 40% and 60%, between 40% and 70%, between 40% and 80%, between 40% and 90%, between 40% and 100%, between 45% and 50%, between 45% and 60%, between 45% and 70%, between 45% and 80%, between 45% and 90%, between 45% and 100%, between 50% and 60%, between 50% and 70%, between 50% and 80%, between 50% and 90%, between 50% and 100%, between 60% and 70%, between 60% and 80%, between 60% and 90%, between 60% and 100%, between 70% and 80%, between 70% and 90%, between 70% and 100%, between 80% and 90%, between 80% and 100%, or between 90% and 100%.
Provided herein are methods of using TrkA polypeptides and polynucleotides for detecting, diagnosing, and monitoring NGF-induced disorders. Provided herein are methods of determining whether a companion animal will respond to TrkA polypeptide therapy. In some embodiments, the method comprises detecting whether the animal has cells expressing NGF using the TrkA polypeptide. In some embodiments, the detection method comprises contacting the sample with an antibody, polypeptide, or polynucleotide and determining whether the level of binding is different from the level of binding of a reference or control sample (e.g., a control). In some embodiments, the methods can be used to determine whether a TrkA polypeptide described herein is an appropriate treatment for a subject animal.
In some embodiments, the sample is a biological sample. The term "biological sample" means a quantity of a substance derived from a living or previously living thing. In some embodiments, the biological sample is a cell or cell/tissue lysate. In some embodiments, biological samples include, but are not limited to, blood (e.g., whole blood), plasma, serum, urine, synovial fluid, and epithelial cells.
In some embodiments, the cells or cell/tissue lysates are contacted with TrkA polypeptide, and binding between the TrkA polypeptide and the cells is determined. When a test cell exhibits binding activity compared to a reference cell of the same tissue type, it may indicate that the subject will benefit from treatment with the TrkA polypeptide. In some embodiments, the test cell is from a tissue of a companion animal.
Various methods known in the art for detecting specific antibody-antigen binding can be used. Exemplary immunoassays that can be performed include Fluorescence Polarization Immunoassay (FPIA), Fluorescence Immunoassay (FIA), Enzyme Immunoassay (EIA), turbidimetric inhibition immunoassay (NIA), enzyme-linked immunosorbent assay (ELISA), and Radioimmunoassay (RIA). Indicator moieties or labeling groups can be attached to the subject antibodies and selected to meet the needs of various applications of the methods, typically determined by the availability of assay devices and compatible immunoassay procedures. Suitable labels include, but are not limited to, radionuclides (e.g., radionuclides)125I、131I、35S、3H or32P), an enzyme (e.g., alkaline phosphatase, horseradish peroxidase, luciferase, or P-lactosidase), a fluorescent moiety or protein (e.g., fluorescein, rhodamine, phycoerythrin, GFP, or BFP), or a luminescent moiety (e.g., Qdot provided by Quantum Dot Corporation, palo alto, ca)TMNanoparticles). General techniques for performing the various immunoassays described above are known to those of ordinary skill in the art.
For diagnostic purposes, the TrkA polypeptide may be labeled with a detectable moiety including, but not limited to, a radioisotope, a fluorescent label, and various enzyme-substrate labels known in the art. Methods of conjugating labels to polypeptides are known in the art. In some embodiments, the TrkA polypeptide need not be labeled, and its presence may be detected, for example, using an antibody that binds to the TrkA polypeptide. In some embodiments, the TRKA polypeptide may be used in any known assay method, such as competitive binding assays, direct and indirect sandwich assays, and immunoprecipitation assays. Zola, Monoclonal antibodyies A Manual of Techniques, pp.147-. anti-NGF antibodies and polypeptides can also be used in vivo diagnostic assays, such as in vivo imaging. Usually with radionuclides (e.g. of the type111In、99Tc、14C、131I、125I、3H or any other radionuclide label (including those outlined herein)) to label the antibody or polypeptide so that the desired cell or tissue can be located using immunoscintigraphy. The TRKA polypeptide can also be used as a staining agent in pathology using techniques well known in the art.
In some embodiments, the TrkA polypeptide is used as a diagnostic agent and the TrkA polypeptide is used as a therapeutic agent. In some embodiments, the first TrkA polypeptide and the second TrkA polypeptide are different.
The following examples illustrate specific aspects of the disclosure and are not intended to limit the disclosure in any way.
Examples
Example 1
Extracellular domain of TrkA
The extracellular domains of canine, feline, and equine TrkA responsible for binding canine, feline, and equine NGF were identified and defined as boundaries. The full-length extracellular domains (ECD v1) of the canine TrkA (SEQ ID NO:2), feline TrkA (SEQ ID NO:7) and equine TrkA (SEQ ID NO:12) were identified from the corresponding full-length polypeptide sequences (SEQ ID NO:1, SEQ ID NO:6 and SEQ ID NO:11, respectively). Exemplary truncated extracellular domain polypeptides of canine TrkA, feline TrkA, and equine TrkA (exemplary ECD v2, v3, and v4) that are presumed to retain NGF binding were identified (e.g., SEQ ID NOS: 3, 4, and 5 (canine TrkA ECD v2, v3, and v 4); SEQ ID NOS: 8, 9, and 10 (feline TrkA ECD v2, v3, and v 4); and SEQ ID NOS: 13, 14, and 15 (equine TrkA ECD v2, v3, and v 4).
Example 2
Design, expression, purification and stability of TrkA ECD polypeptides from CHO cells
To enhance in vivo half-life, NGF binding affinity and purification, exemplary TrkA ECD was fused to an IgG Fc polypeptide bridged with a linker. Preferably, the IgG Fc polypeptide can bind to protein a.
The nucleotide sequences encoding the signal sequence, the canine TrkA ECD v2(SEQ ID NO:3) or canine TrkA ECD v3(SEQ ID NO:4), the linker, and the wild-type canine IgG-B Fc polypeptide were designed, chemically synthesized, and inserted into an expression vector suitable for transfection into CHO host cells. In addition, individual nucleotide sequences encoding the following were similarly designed, synthesized, and cloned into expression vectors: (1) a signal sequence, a cat class TrkA ECD v2(SEQ ID NO:8) or a cat class TrkA ECD v3(SEQ ID NO:9), a linker and a wild type cat class IgG-2Fc polypeptide; and (2) signal sequences, equine TrkA ECD v2(SEQ ID NO:13) or equine TrkA ECD v3(SEQ ID NO:14) and variant equine IgG-2Fc polypeptides (SEQ ID NO: 104; protein A +, C1q-, CD 16-). After transfection into CHO cells and culture, the fusion proteins (SEQ ID NOS: 16, 17, 19, 20, 22 and 23, respectively) were purified from the culture medium by single-step protein A column chromatography.
Differential scanning fluorimetry was used to evaluate the stability of TrkA ECD v2 and TrkA ECD v3 at different pH as reflected by the mean melting point temperature (table 6 below). In this example, the equine TrkA ECD v2 and the equine TrkA ECD v3 fusion polypeptides (SEQ ID NO:22 and SEQ ID NO:23) prepared above were compared. The equine TrkA ECD v3(SEQ ID NO:23) was slightly more stable than the equine TrkA ECD v2(SEQ ID NO:22) in terms of thermostability at the tested pH.
Table 6.
Example 3
Demonstration of NGF binding Activity
The binding activity of the canine, feline, and equine TrkA ECD v2 and v3 fusion polypeptides (SEQ ID NOS: 16, 17, 19, 20, 22, and 23) prepared in example 2 with commercially available human NGF was considered. The following binding assay was performed using the biosensor Octet. Briefly, human NGF (Sino Biological, Inc.; Cat. No. 11050-HNAC) was biotinylated. Free unreacted biotin was removed from biotinylated NGF by extensive dialysis. Biotinylated NGF was captured on streptavidin sensor tips. Monitor forNGF association with TrkA-Fc (25ug/mL) was measured for 600 seconds. Dissociation was monitored for 600 seconds. Buffer only blank curves were subtracted to correct for any drift. Using ForteBioTMData analysis software fitting data to 1:1 binding model to determine kon、koffAnd Kd. The buffers used for the dilution and all binding steps were: 20mM phosphate, 150mM NaCl, pH 7.2.
Figures 1 and 2 are sensorgrams showing that both canine, feline and equine TrkA ECD v2 and v3 fusion polypeptides bind NGF. Figure 1 shows a sensorgram comparing the binding affinities of canine and feline TrkA v2 and v3 to NGF. Irrelevant canine and feline IgG-Fc fusion proteins were used as negative controls. Figure 2 shows a sensorgram comparing the binding affinity of equine TrkA v2 and v3 for NGF. Of the three species, all TrkA ECD and NGF had Kd at 1X10-9And M is in the range.
Example 4
Cell functional Activity of TrkA ECD-IgG Fc Polypeptides
TF1 cells (ATCC accession number CRL-2003), a human erythroleukemia cell line that expresses NGF receptors on the cell surface, were used in proliferation assays. Recombinant human NGF stimulates cell proliferation of TF-1 cells in the absence of other essential growth factors (e.g., erythropoietin, IL3, or GM-CSF). See Kitamura, T. et al, "assessment and characterization of a unique human cell lines and promoters on GM-CSF, IL-3, or erythropoeitin," J.cell Physiol.140(2):323-34 (1989); com/pro ucts/recombiant-human-beta-ngf-protein-256-gf # products-details. Cells were grown in RP MI1640(Gibco, Cat. No. 11875) supplemented with 10% heat-inactivated fetal bovine serum (Sigma, Cat. No. 2868) and 2nM/ml human GM-CSF (R & D System, Cat. No. 215-GM-010). Cells at exponential growth phase were washed with PBS, resuspended in the above medium without GM-C SF, and plated in 96-well plates. The equine TrkA ECD v 2-variant equine IgG2Fc (SEQ ID NO: 22; protein A +, C1q-, CD16-) was added in a series of dilutions, followed by addition of NGF at 10ng/ml (Sino Biological, Inc.; catalog No. 11050-HNAC). Irrelevant monoclonal antibodies were used as negative controls.
Cells were incubated at 37 ℃ in a total volume of 100. mu.l with 5% CO2And (4) incubating for 48 hours. At the end of the incubation, the cells were cooled at room temperature and were incubated by using CellTiter-Proliferation/viability was analyzed by luminometric cell viability assay (Promega, catalog No. G7570) which measures cellular ATP content. In this assay, 100 μ l of pre-mixed reagents a and B were added to each well. After shaking on an orbital shaker for about 2 minutes, the cells were lysed. In the presence of Mg2+ and ATP present in the cells, monooxygenation of luciferin is catalyzed by luciferase, producing a luminescent signal proportional to the amount of ATP in the cells. The amount of ATP is proportional to the number of cells present in the culture. Plates were incubated at room temperature for 10min to stabilize the luminescence signal and luminescence was detected using a Synergy HT microplate reader (Biotek, knonski, budd). Data were analyzed using a 4-parameter logistic fit and it was demonstrated that the TrkA ECD-IgG Fc polypeptide neutralized NGF activity in this TF1 cell proliferation assay. The IC50 for the TrkA ECD-Fc polypeptide was 5.4 nM. See fig. 3.
Example 5
Engineering N-linked glycosylation of TrkA ECD polypeptides
To further enhance the pharmacokinetics of TrkA ECD polypeptides, additional N-linked glycosylation sites may be introduced into wild-type canine TrkA, feline TrkA, and equine TrkA. For example, one, two, three or four additional N-linked glycosylation sites can be introduced into the TrkA ECD amino acid sequence at non-overlapping positions. The N-linked glycosylation site can have a consensus sequence Asn-Xaa-Ser/Thr, wherein Xaa is any amino acid except proline. The addition of one or more glycosylation sites can increase the molecular size of the TrkA ECD polypeptide, provide more sialylation sites, and/or improve the half-life of the polypeptide in the serum of an animal.
The TrkA ECD polypeptide may be produced by a mammalian cell under conditions that enhance sialylation. In addition, the TrkA ECD polypeptide may be conjugated by an amine or further pegylated or polysialylated on a glycan. For example, chemical polysialylation can be introduced into glycosylation sites.
Table 7 lists amino acid substitutions of the canine TrkA ECD v2, v3, and v4, which may be used to create one or more additional N-linked glycosylation sites.
Table 7.
X indicates any amino acid other than proline (e.g. E, V, A, I etc.).
Table 8 lists amino acid substitutions of cat TrkA ECD v2, v3, and v4, which can be used to create one or more additional N-linked glycosylation sites.
Table 8.
X indicates any amino acid other than proline (e.g. E, V, A, I etc.).
Table 9 lists the amino acid substitutions of the equine TrkA ECD v2, v3, and v4 that may be used to create one or more additional N-linked glycosylation sites.
Table 9.
X indicates any amino acid other than proline (e.g. E, V, A, I etc.).
Example 6
Intramolecular TrkA ECD disulfide
To increase the stability of TrkA ECD polypeptides, suitable sites for additional intramolecular disulfide binding were identified by three-dimensional protein modeling and analysis. Additional disulfide binding may prevent the TrkA ECD polypeptide from unfolding and enhance protease resistance, resulting in enhanced product shelf-life stability and enhanced pharmacokinetics in vivo. For example, cysteine residues can be incorporated into the canine, feline, or equine TrkA ECD V2 or V3 (V7C and A89C (for canine or feline sequences) or D89C (for equine sequences)) at amino acid positions 7 and 89 of SEQ ID NO 3, 4, 8, 9, 13, or 14. Cysteine residues may also be incorporated into the canine, feline, or equine TrkA ECD V4 (V5C and a87C (for canine or feline sequences) or D87C (for equine sequences)) at amino acid positions 5 and 87 of SEQ ID NOs 5, 10, or 15. Exemplary TrkA ECD polypeptides with additional disulfide pairs include SEQ ID NOs 25, 26, 27, 28, 29, 30, 31, 32, and 33.
Example 7
Exemplary contiguous polypeptides comprising TrkA ECD and IgG Fc
Contiguous polypeptides comprising a canine, feline, or equine TrkA ECD v1, v2, v3, or v4 polypeptide (e.g., SEQ ID NO:2, 3, 4, 5, 7, 8, 9, 10, 12, 13, 14, 15, 25, 26, 27, 28, 29, 30, 31, 32, or 33) and a wild-type or variant IgG Fc polypeptide of the corresponding companion animal can be designed, expressed, and purified for characterization. Preferably, TrkA ECD linked to an IgG Fc polypeptide having protein a binding, reduced or non-measurable binding to C1q (e.g., to reduce CDC function), and/or reduced or non-measurable binding to CD16 (e.g., to reduce ADCC function) is used for administration to companion animals suffering from NGF-induced pain. Exemplary wild-type canine, feline, and equine IgG Fc polypeptides comprise the amino acid sequences of SEQ ID NOS 33-39, 70-77, or 86-90. Exemplary variant canine, feline, and equine IgG Fc polypeptides comprise amino acid sequences such as those described in examples 8-11 (e.g., SEQ ID NOs: 40-69, 78-85, or 91-109).
Continuous polypeptides comprising at least one TrkA ECD polypeptide (e.g., ECD v1, v2, v3, and/or v4) and a wild-type or variant canine, feline, or equine IgG Fc polypeptide described herein can be prepared based on the following formula:
formula (I): TrkA ECD 1-L1-Fc;
formula (II): Fc-L1-TrkA ECD 1;
formula (III): TrkA ECD 1-L1-Fc-L2-TrkA ECD 2;
formula (IV): TrkA ECD 1-L1-TrkA ECD 2-L2-Fc; or
Formula (V): Fc-L1-TrkA ECD 1-L2-TrkA ECD 2,
wherein TrkA ECD 1 is a first TrkA ECD polypeptide, TrkA ECD 2 is a second TrkA ECD polypeptide (e.g., the same TrkA ECD polypeptide or a different TrkA ECD polypeptide), L1 and L2 are optional linkers; and Fc is a wild-type or variant IgG Fc polypeptide of the companion animal species. Optionally, the contiguous polypeptide comprises a signal sequence. Exemplary constructs of formulae I-V may comprise a third, fourth or fifth, etc. TrkA ECD after or before any of TrkA ECD 1 or TrkA ECD 2. The third, fourth or fifth, etc. TrkA ECD may be the same TrkA ECD polypeptide as TrkA ECD 1 or TrkA ECD 2 or a different TrkA ECD polypeptide.
For example, the contiguous polypeptide may comprise at least one canine TrkA ECD polypeptide (e.g., SEQ ID NO:2, 3, 4, 5, 25, 26, or 27) and wild-type canine IgG polypeptide (e.g., SEQ ID NO:34, 35, 36, 37, 38, or 39), a variant canine IgG-A Fc polypeptide (e.g., SEQ ID NO:40, 43, 199, or 200), a variant canine IgG-B Fc polypeptide (e.g., SEQ ID NO:46, 48, 49, 50, 51, 52, 53, 54, 55, 64, 65, 66, 67, 197, 198, 203, 204, 205, 206, 207, 208, 209, or 210), a variant canine IgG-C Fc polypeptide (e.g., SEQ ID NO:41, 44, 47, 56, 57, 58, 59, 60, 61, 52, 63, 68, or 69), or a variant IgG-DFc polypeptide (e.g., SEQ ID NO:42, 45, 201, or 202), as described herein.
The contiguous polypeptide can comprise at least one cat TrkA ECD polypeptide (e.g., SEQ ID NOs: 7, 8, 9, 10, 28, 29, or 30) and a wild-type cat IgG Fc polypeptide (e.g., 86, 87, 88, 89, or 90), a variant cat IgG1a Fc polypeptide (e.g., SEQ ID NOs: 91, 92, 96, or 97), a variant cat IgG1b Fc polypeptide (e.g., SEQ ID NOs: 93, 94, 98, or 99), or a variant cat IgG2Fc polypeptide (e.g., SEQ ID NOs: 95, 100, or 107), as described herein.
The contiguous polypeptide may comprise at least one equine TrkA ECD polypeptide (e.g., SEQ ID NO:12, 13, 14, 15, 31, 32, or 33) and a wild-type equine IgG Fc polypeptide (e.g., SEQ ID NO:70, 71, 72, 73, 74, 75, 76, or 77), a variant equine IgG1Fc polypeptide (e.g., SEQ ID NO:82), a variant equine IgG2Fc polypeptide (e.g., SEQ ID NO:78, 79, 101, 102, 103, 104, 105, 106, 108, or 109), a variant equine IgG3 Fc polypeptide (e.g., SEQ ID NO:83), a variant equine IgG4 Fc polypeptide (e.g., SEQ ID NO:84), a variant equine IgG5 Fc polypeptide (e.g., SEQ ID NO:80), a variant equine IgG6Fc polypeptide (e.g., SEQ ID NO:81), or a variant equine IgG7Fc polypeptide (e.g., SEQ ID NO: 85).
The linker may be a flexible non-structural linker, such as a glycine and/or serine rich linker. A flexible extension may be added to the C-terminus of the contiguous polypeptide. The extension may comprise one, two, three, four, five, six, seven, eight or more glycine residues.
The contiguous polypeptide comprising a TrkA ECD may further comprise at least one ECD of a NGFR polypeptide, such as SEQ ID NO 135, 137 or 139.
Protein A columns can be used (Protein a affinity resin, Repligen) affinity purifying a continuous polypeptide comprising an Fc IgG polypeptide having protein a binding. The continuous polypeptide may also be isolated via other chromatographic methods such as ion exchange column chromatography, hydrophobic interaction column chromatography, mixed mode column chromatography (e.g., CHT), or multimodal column chromatography (e.g., captomamc). Low pH or other virus inactivation and virus removal steps may be applied. The purified protein may be mixed with excipients and sterilized by filtration to prepare the pharmaceutical composition of the present invention.
Dimerization, aggregation and/or presence of sulfide linkages of the resulting proteins can be assessed by HPLC gel filtration and/or SDS-PAGE analysis in the absence and presence of a reducing agent (DTT).
Example 8
For increased protein A binding and/or decreased complement fixation
And/or reduced CD16 binding variant canine IgG Fc polypeptides
Purification of antibodies using protein a affinity is a mature process. However, of the four subtypes of canine IgG, only IgG-B Fc (e.g., SEQ ID NO:35 or SEQ ID NO:36) has protein A binding affinity. Canine IgG-A Fc (e.g., SEQ ID NO:34), IgG-C Fc (e.g., SEQ ID NO:37 or SEQ ID NO:38) and IgG-D Fc (e.g., SEQ ID NO:39) have weak or NO measurable protein A binding affinity. Variant canine IgG-A Fc, IgG-C Fc, and IgG-D Fc polypeptides were designed for altered protein A binding.
Furthermore, canine IgG-B Fc and IgG-C Fc have complement activity and bind to C1q, while canine IgG-a Fc and IgG-D Fc have weak or no measurable binding affinity for C1 q. To potentially reduce C1q binding and/or to potentially reduce complement-mediated immune responses, variant canine IgG-B Fc and IgG-C Fc polypeptides were designed.
Furthermore, the canine IgG-B Fc and IgG-C Fc have CD16 binding activity. To potentially reduce binding of CD16 to IgG-B Fc and IgG-C Fc, and/or to potentially reduce ADCC, variant canine IgG-B Fc and IgG-C Fc polypeptides were designed.
Table 10 below summarizes the binding characteristics of the proteins A, C1q and CD16 for the canine IgG Fc subtype. Notably, none of the wild-type canine IgG Fc subtypes bind protein a and lack C1q binding and/or CD16 binding.
Table 10.
(-) indicates low or no measurable binding activity.
Using three-dimensional protein modeling and protein sequence analysis, the sequences of canine IgG-B Fc that are likely to be in contact with protein A were identified. Two approaches were used to design variant canine IgG-A, IgG-C and IgG-D Fc polypeptides for increased protein a binding. For the first approach, variant canine IgG-A, IgG-C and IgG-D Fc polypeptides were designed to have the same protein A binding motif sequence as canine IgG-B Fc (e.g., SEQ ID NO:40, SEQ ID NO:41, and SEQ ID NO:42, respectively). For the second approach, variant canine IgG-A Fc I (21) T/Q (207) H (SEQ ID NO:43), variant canine IgG-C Fc I (21) T (SEQ ID NO:44), and variant canine IgG-D Fc I (21) T/Q (207) H (SEQ ID NO:45) were designed to have one or two amino acid substitutions in the protein A binding region to correspond to a canine IgG-B Fc sequence.
In addition, variant canine IgG-A Fc, IgG-C Fc, and IgG-D Fc polypeptides having increased protein A binding can be prepared having one or more amino acid substitutions set forth in Table 11.
Table 11.
The amino acid positions listed are relative to the indicated SEQ ID no.
To potentially reduce binding of C1q to canine IgG-B Fc and IgG-C Fc, and/or to potentially reduce a complement-mediated immune response, variant canine IgG-B Fc and IgG-C Fc polypeptides having an amino acid substitution of Lys with any amino acid other than Lys at the amino acid position corresponding to position 93 of SEQ ID NO:35 or SEQ ID NO:37, respectively, can be prepared. These amino acid substitutions were identified following protein sequence analysis and 3-D structural modeling of canine IgG-B Fc and IgG-C Fc in comparison to canine IgG-A Fc and IgG-D Fc (which are known to exhibit no complement activity). For example, variant canine IgG-B Fc K (93) R (SEQ ID NO:46) and variant canine IgG-C Fc K (93) R (SEQ ID NO:47) can be prepared. Reduced binding between human C1q and fusion proteins comprising variant canine IgG-B Fc K (93) R was observed compared to fusion proteins comprising wild-type canine IgG-B Fc.
To potentially reduce binding of CD16 to IgG-B Fc and IgG-C Fc, and/or to potentially reduce ADCC, variant canine IgG-B Fc and IgG-C Fc polypeptides having one or more amino acid substitutions set forth in Table 12 can be prepared (e.g., SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, and/or SEQ ID NO: 62). The one or more amino acid substitutions were identified following protein sequence analysis and 3-D structural modeling of canine IgG-B and IgG-C compared to IgG-A and IgG-D (with the understanding that they do not exhibit ADCC activity).
Table 12.
The amino acid positions listed are relative to the indicated SEQ ID no.
Since wild-type canine IgG-C Fc lacks protein a binding and has C1q binding, a dual variant canine IgG-C Fc that binds protein a and has reduced binding to C1q (e.g., SEQ ID NO:63) can be prepared by combining one or more of the amino acid substitutions listed in table 11 with a K (93) R substitution or a K (93) X substitution (where X is any amino acid except Lys). Double variant canine IgG-B Fc or double variant canine IgG-C Fc (e.g., SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, and/or SEQ ID NO:69) with reduced binding to C1q and reduced binding to CD16 can be prepared by combining one or more of the amino acid substitutions listed in table 12 with a K (93) R substitution or a K (93) R substitution (where X is any amino acid other than Lys). A triple variant canine IgG-C Fc that binds to protein a and has reduced binding to C1q and CD16 can be prepared by combining one or more amino acid substitutions listed in table 11 and one or more amino acid substitutions listed in table 12 with a K (93) R substitution or a K (93) X substitution (where X is any amino acid except Lys).
Binding of any variant canine IgG Fc to protein A, CD16 and/or C1q can be determined and compared to binding of another IgG Fc (e.g., a corresponding wild-type canine IgG Fc, another wild-type or variant canine IgG Fc, or a wild-type or variant IgG Fc from another companion animal, etc.) to protein A, CD16 and/or C1 q.
Binding assays can be performed using Octet biosensors. Briefly, target molecules (e.g., protein A, C1q, CD16, etc.) can be biotinylated and free-flowing removedThe separated unreacted biotin (e.g., by dialysis). The biotinylated target molecules are captured on streptavidin sensor tips. The association of target molecules with various concentrations (e.g., 10 μ g/mL) of IgG Fc polypeptides was monitored for a specified time or until steady state was reached. Dissociation was monitored for a specified time or until steady state was reached. A blank curve of buffer only can be subtracted to correct for any drift. Using ForteBioTMData analysis software fitting data to 1:1 binding model to determine kon、koffAnd Kd。
Example 9
For increased protein A binding
And/or reduced complement fixation variant equine IgG Fc polypeptides
Among the seven subtypes of equine IgG, IgG1Fc (e.g., SEQ ID NO:70), IgG3 Fc (e.g., SEQ ID NO:73), IgG4 Fc (e.g., SEQ ID NO:74), IgG7Fc (e.g., SEQ ID NO:77) have protein A binding affinity. Equine IgG2Fc (e.g., SEQ ID NO:71 or SEQ ID NO:72), IgG5 Fc (e.g., SEQ ID NO:75), and IgG6Fc (e.g., SEQ ID NO:76) have weak or NO measurable protein A binding affinity. Variant equine IgG2Fc, IgG5 Fc, and IgG6Fc polypeptides were designed for use in altering protein a binding.
Furthermore, equine classes IgG2Fc, IgG5 Fc and IgG6Fc have weak or no measurable binding affinity for C1q, while equine classes IgG1Fc, IgG3 Fc, IgG4 Fc and IgG7Fc bind to C1 q. To potentially reduce C1q binding and/or to potentially reduce complement-mediated immune responses, variant equine IgG1Fc, IgG3 Fc, IgG4 Fc, and IgG7Fc polypeptides were designed.
Table 13 below summarizes the protein a and C1q binding characteristics of the equine IgG Fc subtype. Notably, none of the wild-type equine IgG Fc subtypes lack C1q binding and bind protein a.
Table 13.
(-) indicates low or no measurable binding activity.
Using three-dimensional protein modeling and protein sequence analysis, sequences of equine IgG1Fc, IgG3 Fc, IgG4 Fc, and IgG7Fc that are likely to contact protein a were identified. Variant equine IgG2Fc, IgG5 Fc, and IgG6Fc polypeptides with increased protein a binding can be prepared having one or more amino acid substitutions listed in table 14.
Table 14.
Listed amino acid positions are relative to the indicated SEQ ID NO.
For example, variant equine IgG2Fc, IgG5 Fc, and IgG6Fc polypeptides are designed to have one or more amino acid substitutions in the protein a binding region to correspond to the amino acid sequence of a wild-type equine IgG Fc that binds protein a. Variant equine IgG2Fc F (203) Y (SEQ ID NO:78) with increased protein A binding can be prepared; variant equine IgG2Fc A (15) T/F (203) Y (SEQ ID NO: 79); variant equine IgG5 Fc V (199) L/E (200) Y (SEQ ID NO: 80); and variant equine IgG6Fc I (199) L/R (200) H/H (201) N/T (202) H (SEQ ID NO: 81).
To potentially reduce binding of C1q to equine IgG1Fc, IgG3 Fc, IgG4 Fc, and IgG7Fc, and/or to potentially reduce a complement-mediated immune response, variant canine IgG1Fc, IgG3 Fc, IgG4 Fc, and IgG7Fc polypeptides can be prepared that have an amino acid substitution of Lys with any amino acid other than Lys at an amino acid position corresponding to position 87 of SEQ ID NO:70, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:77, respectively. These amino acid substitutions were identified following protein sequence analysis and 3-D structural modeling of equine classes IgG1Fc, IgG3 Fc, IgG4 Fc, and IgG7Fc in comparison to equine classes IgG2Fc, IgG5 Fc, and IgG6Fc (which are known to exhibit no complement activity). For example, variant equine IgG1Fc K (87) S (SEQ ID NO:82), variant equine IgG3 Fc K (87) S (SEQ ID NO:83), variant equine IgG4 Fc K (87) S (SEQ ID NO:84), and variant equine IgG7Fc K (87) S (SEQ ID NO:85) can be prepared.
Binding of any variant equine IgG Fc to protein a and/or C1q can be determined and compared to binding of another IgG Fc (e.g., a corresponding wild-type equine IgG Fc, another wild-type or variant equine IgG Fc, or a wild-type or variant IgG Fc from another companion animal, etc.) to protein a and/or C1 q. The binding assay described in example 8 can be used.
Example 10
Variant cat class IgG Fc polypeptides for reduced complement fixation
Each of the three subtypes of feline IgG, IgG1a Fc (SEQ ID NO:86 or SEQ ID NO:87), IgG1b Fc (SEQ ID NO:88 or SEQ ID NO:89), and IgG2Fc (SEQ ID NO:90) has protein A binding affinity. However, only cat class IgG2Fc has weak or no measurable binding affinity for C1q, while cat classes IgG1a Fc, IgG1b Fc bind to C1 q. To potentially reduce C1q binding and/or to potentially reduce complement-mediated immune responses, variant feline IgG1a Fc and IgG1b Fc polypeptides were designed.
Table 15 below summarizes the protein a and C1q binding characteristics of cat IgG Fc subtypes. Notably, none of the wild-type equine IgG Fc subtypes lack C1q binding and bind protein a.
Table 15.
(-) indicates low or no measurable binding activity.
To potentially reduce binding of C1q to feline IgG1a Fc and IgG1b Fc, and/or to potentially reduce complement-mediated immune responses, variant feline IgG1a Fc and IgG1b Fc polypeptides can be prepared that have an amino acid substitution of Pro with any amino acid other than Pro at an amino acid position corresponding to position 198 of SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, or SEQ ID NO: 89. These amino acid substitutions were identified following protein sequence analysis and 3-D structural modeling of cat class IgG1a Fc and IgG1b Fc compared to cat class IgG2Fc (which is understood to exhibit no complement activity). For example, variant cat class IgG1a Fc P (198) A (e.g., SEQ ID NO:91 or SEQ ID NO:92) and variant cat class IgG1b Fc P (198) A (e.g., SEQ ID NO:93 or SEQ ID NO:94) can be prepared.
Binding of any variant cat class IgG Fc to C1q can be determined and compared to binding of another IgG Fc (e.g., a corresponding wild-type cat class IgG Fc, another wild-type or variant cat class IgG Fc, or a wild-type or variant IgG Fc of another companion animal, etc.) to C1 q. The binding assay described in example 8 can be used.
Example 11
Variant IgG Fc polypeptides for increasing and/or enhancing disulfide formation
Three-dimensional protein modeling analysis of several orthologous hinge structures was used to determine the approximate location for modifying the feline IgG2 hinge to increase disulfide formation. To increase disulfide formation at the hinge of cat IgG2, the hinge sequence can be modified by substituting amino acids with cysteines. For example, a variant cat class IgG2Fc (SEQ ID NO:95) with a modified hinge can be prepared by substituting a cysteine for a glycine at the amino acid position corresponding to position 14 of SEQ ID NO: 90. Other variant feline IgG2Fc polypeptides can be prepared having a modified hinge comprising a cysteine at an amino acid position corresponding to position 8, position 9, position 10, position 11, position 12, position 13, position 15, or position 16 of SEQ ID No. 90.
Cat and horse IgG hinges were modified to enhance disulfide formation using additional three-dimensional protein modeling analysis of several orthologous hinge structures. To enhance disulfide formation at the hinge of cat class IgG, the hinge sequence can be modified by: proline is substituted for lysine (e.g., K16P) at a position corresponding to position 16 of wild-type or variant cat class IgG1a (e.g., SEQ ID NO:86 or SEQ ID NO:87), cat class IgG1b (e.g., SEQ ID NO:88 or SEQ ID NO:89), or cat class IgG2 (e.g., SEQ ID NO: 90). Examples of amino acid sequences of variant feline IgG polypeptides having modified hinges include SEQ ID NO 96, SEQ ID NO 97, SEQ ID NO 98, SEQ ID NO 99, and SEQ ID NO 100.
To enhance disulfide formation at the equine IgG hinge, the hinge sequence can be modified by: the substitution of cysteine with serine at a position corresponding to position 3 of a wild-type or variant equine IgG having a hinge (e.g., IgG2Fc (SEQ ID NO:72)) and/or the substitution of glutamine with proline at a position corresponding to position 20 of an equine IgG having a hinge (e.g., IgG2Fc (SEQ ID NO:72)) for glutamine (e.g., C3S and/or Q20P examples of amino acid sequences of variant equine IgG polypeptides having modified hinges include SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:103, SEQ ID NO:104, SEQ ID NO:105, and SEQ ID NO: 106.
The above amino acid substitutions may be incorporated into the hinge of a wild-type or variant Fc polypeptide described herein.
Three-dimensional protein modeling was used to design cat and horse variant IgG Fc polypeptides comprising sequences from the hinge region of different IgG isotypes for enhanced recombinant production and improved hinge disulfide formation. Variant feline IgG2Fc polypeptides (e.g., SEQ ID NO:107) can be prepared that comprise a sequence from the hinge region of a feline IgG1a or IgG1 b. In addition, variant equine IgG2Fc polypeptides (e.g., SEQ ID NO:108 and SEQ ID NO:109) comprising sequences from the hinge region of equine IgG1 can be prepared.
The level of recombinant production of the variant IgG Fc polypeptide and/or the level of hinge disulfide formation can be determined and compared to that of another IgG Fc (e.g., a corresponding wild-type IgG Fc of the same or different isotype, or a wild-type or variant IgG Fc of another companion animal) by SDS-PAGE analysis under reducing and non-reducing conditions.
Example 12
TrkA-Fc buffer formulation
The thermal stability of the canine TrkA-Fc was analyzed in various buffer formulations. Buffers containing 20mM sodium phosphate (pH 6.2, 6.6, 7.2 and 7.6), 20mM sodium citrate (pH 4.4, 5.2, 5.8 and 6.4), 20mM histidine (pH 5.5, 6.0 and 6.5) and 20mM sodium acetate (pH 5.2) are contemplated. Sodium chloride was used in all buffers at final concentrations of 40mM or 140 mM. The melting temperatures (Tm 1 and Tm 2) of exemplary canine TrkA-Fc polypeptide (SEQ ID NO:126) at a concentration of 6. mu.g/. mu.L in each buffer were measured in duplicate by differential scanning fluorescence techniques from 20 ℃ to 95 ℃. Table 16 lists the mean Tm values for the canine TrkA in the various buffers tested.
Table 16.
Results from one sample
Formulations 6A, 7B, 9A, and 12B containing lower concentrations of sodium citrate, histidine, or sodium acetate buffers and pH between 5 and 6 may be more desirable for TrkA ECD fusion polypeptides.
Formulations 6A, 7B, 9A, 12B and PBS (pH 7.2) were used to study the stability of samples of exemplary canine TrkA-Fc polypeptide (SEQ ID NO:126) stored under stress at 45 ℃ for 3 days. The aggregation status of the five samples was evaluated by HPLC gel filtration analysis and no significant aggregation was identified. Five samples were then stored at 55 ℃ for another day. Significant aggregation was observed in samples stored in PBS (pH 7.2), but not in samples stored in formulations 6A, 7B, 9A, and 12B.
Example 13
In vivo reduction of NGF with TrkA-Fc polypeptide
NGF levels are elevated in synovial fluid of human patients with various types of chronic arthritis. See Aloe L et al, "New growth factor in the synthetic fluid of tissues with chronic Arthritis," Arthritis Rheum.1992,35(3): 351-5. The effect of the canine TrkA-Fc polypeptide on NGF levels was tested in a rat MIA-induced osteoarthritis model.
The study protocol was reviewed and approved by the Institutional Animal Ethics Committee (IAEC) (approval No.: B-070). Male Sprague-Dawley (Sprague Dawley) rats were anesthetized with isoflurane and given a single intra-articular injection of 3mg monosodium iodoacetate (Sigma, cat # I2512, st. louis, missouri) through the infrapatellar ligament of the right knee (osteoarthritis). MIA was dissolved in saline and administered in a volume of 50 μ l using a 30 gauge, 0.5 inch needle. See Combe R et al, "The monosodious iodide model of osteoarthritis a model of cyclic nociceptive pain in rates? Neurosci Lett.2004,370(2-3): 236-40. Control rats were injected with an equal volume of saline and allowed to recover. After recovery from the procedure, animals were returned to the cages in groups of three, using a 12h light/dark cycle and ad libitum access to food and water.
Weekly administrations of the canine TrkA-Fc polypeptides (1mg/kg (group 3) and 10mg/kg (group 4)) were performed on days 8, 15 and 22 post-MIA injection. Synovial NGF levels were measured on day 28. The treatment groups are summarized in table 17 below.
Table 17.
Knee samples from 3-4 rats from each treatment group were collected after performance and knee diameter measurements were completed. Trimmed samples of soft tissue within the knee (synovium and anterior of the ipsilateral knee lateral capsule) were collected and minced into small pieces of approximately 2mm in size, rinsed well in PBS, and frozen in liquid nitrogen until use. Tissue samples were crushed to powder and treated with protease inhibitor (clomplete)TM200- μ l lysis reagent (CelLytic) of EDTA-free protease inhibitor cocktail, Cat No. 4693132001, Sigma-Aldrich)TMMT cell lysis reagent (C3228), Sigma-Aldrich). The samples were centrifuged at 14,000rpm for 10min at 4 ℃, and then the supernatant was collected for assay. The concentration of NGF was measured using an ELISA kit (Millipore, Temankura, Calif.). Use of BCA proteinThe assay kit (catalog No. BCA1-1KT, Sigma, usa) measures the total protein concentration in all samples.
Figure 4 shows the concentration of NGF in the synovium of each of the four animal groups. The MIA-induced animal groups (groups 3 and 4) receiving the canine TrkA-Fc polypeptide showed a dose-dependent decrease in NGF concentration in the synovium compared to the untreated MIA-induced control group (group 2). For reasons still under investigation, in this experiment, the treated animals did not show a corresponding reduction in pain, as has been expected based on published studies involving the treatment of animals with NGF antibodies and TrkA-IgG fusion molecules. See Ro LS et al, "Effect of NGF and anti-NGF on neuropathic pain in rates of neurological compliance in the future," pain.1999,79(2-3): 265-74; gearing D et al, "A full nitrogenous anti-NGF monomeric anti for pain relief in logs." BMC Vet Res.2013,9:226doi: 10.1186/1746-; gearing D et al, "In vitro and In vivo characteristics of a fully fluorinated therapeutic anti-new facial factor monoclonal antibody for the treatment of the disease In cat, J.vet Intern Med.2016,30(4): 1129-37; McMahon SB et al, "The biological effects of endogenous growth factors on adult sensor neurones modified by a trkA-IgG fusion moleculae," Nat Med.1995,1(8): 774-80.
Example 14
Screening for variant canine IgG-B polypeptides with enhanced binding to canine FcRn/B2M
The canine FcRn (SEQ ID NO:195) and canine B2M (SEQ ID NO:196) heterodimer complexes with poly-His tags were transiently expressed in HEK cells and purified using Ni-NTA chromatography.
A canine IgG-B Fc phage library was rapidly screened for expression, biophysical properties, and affinity (FASEBA). Briefly, the open reading frame for the canine IgG-B Fc polypeptide was subcloned into the plasmid pfasaba. Based on three-dimensional protein modeling of the canine IgG-B/canine FcRn/canine B2M complex, twelve amino acid positions of canine IgG-B were identified as likely to be involved in binding between IgG-B and FcRn/B2M. The twelve positions identified for canine IgG-B are Thr (21), Leu (22), Leu (23), Ile (24), Ala (25), Thr (27), Gly (80), His (81), Gln (82), Leu (85), Met (201), and Asn (207) of SEQ ID NO:35 or SEQ ID NO: 36.
Twelve single-site NNK mutation libraries of canine IgG-B Fc were prepared such that each library would include variant IgG-B Fc polypeptides having each of the 20 possible amino acids substituted at each of the twelve sites. Each phage library was panned against the canine FcRn/B2M complex at pH 6.0. After three rounds of panning, a total of 53 Fc phage clones were identified as likely to have enhanced FcRn/B2M binding, and mutations were identified by sequencing.
A single e.coli (e.coli) colony expressing each of the 53 variant canine IgG-B Fc polypeptides with the SASA tag was cultured and induced to express Fc polypeptides. Cell culture media containing variant canine IgG-B Fc polypeptides were exposed to immobilized BSA on plates or Biacore chips. Plates or chips bound to variant canine IgG-B Fc polypeptides were exposed to the soluble canine FcRn/B2M complex to screen for slow off-rate (koff) at pH6. Each variant IgG-B Fc polypeptide that exhibits a slower koff to canine FcRn/B2M complex compared to a wild-type IgG-B Fc polypeptide was identified. Four lead variant canine IgG-B polypeptides were identified: l (23) Y (SEQ ID NO: 198; "Y00"); l (23) F (SEQ ID NO: 197; "F00"); l (23) M; and L (23) S.
The koff of each lead variant canine IgG-B polypeptide was further investigated. Biotinylated canine FcRn/B2M complex was immobilized on a Biacore chip and exposed to each variant canine IgG-B polypeptide as an analyte using Biacore T200 at pH 6.0. The koff (1/s) of the wild-type canine IgG-B Fc polypeptide was 1.22x10-1(ii) a The koff (1/s) of the variant canine IgG-B Fc polypeptide L (23) Y ("Y00") was 1.38x10-2(ii) a The koff (1/s) of the variant IgG-B Fc polypeptide L (23) F ("F00") was 6.31x10-2And 8.47x10-2(ii) a The koff (1/s) of the variant canine IgG-B polypeptide L (23) M was 1.26x10-1(ii) a And the koff (1/S) of the variant canine IgG-B polypeptide L (23) S is 2.41x10-1。
Binding analysis was performed using Biacore T200. Briefly, leader variant canine IgG-B Fc polypeptides with SASA tags were each immobilized onto S-series sensor chip CM 5. The association of each variant IgG-B Fc polypeptide with various concentrations of the canine FcRn/B2M complex (12.5, 25, 50, 100, and 200nM) was monitored at 25 ℃ until steady state was reached. A running buffer (pH 6.0) of 10mM HEPES, 500mM NaCl, 3mM EDTA, 0.005% Tween-20 was used. Buffer only blank curves were used as controls. The results are presented in fig.5 to 9. The steady state Kd of the wild-type canine IgG-B Fc polypeptide was 1.25x10-6(FIG. 5); the steady state Kd of the variant canine IgG-B Fc polypeptide L (23) Y ("Y00") was 1.13x10-7(FIG. 6); the steady state Kd of the variant canine IgG-B Fc polypeptide L (23) F ("F00") was 3.67x10-7(FIG. 7); and the steady state Kd of the variant canine IgG-B Fc polypeptide L (23) M is 4.06x10-7(FIG. 8); and the steady state Kd of the variant canine IgG-B Fc polypeptide YTE is 8.62x10-8(FIG. 9).
Example 15
Phe mutations in canine IgG enhance canine FcRn interaction
The affinity of the variant canine Fc polypeptides for FcRn was evaluated in the context of chimeric antibodies. Antibody variable light chains fused to canine kappa light chains and variable heavy chains fused to variant canine IgG-A Fc polypeptides comprising SEQ ID NO 199 (F00; protein A +; C1 q-; CD16-) or SEQ ID NO 200 (protein A +; C1q +; CD16+) and to variant canine IgG-D Fc polypeptides comprising SEQ ID NO 201 (F00; protein A +; C1 q-; CD16-) or SEQ ID NO 202 (protein A +; C1q +; CD16+) were expressed.
Binding analysis was performed using the biosensor OctetRed as follows. Briefly, biotinylated TNF α was captured on the streptavidin sensor tip. The association of 20. mu.g/mL of antibody was with TNF α binding. The complexes were then used to bind canine FcRn (50 μ g/mL) at pH 6.0. Dissociation was performed at pH 7.2.
The Phe mutation enhanced canine FcRn binding at low pH (pH6.0, 20mM sodium citrate, 140mM NaCl) as shown by the binding profiles of the chimeric variant canine IgG-a "F00" antibody (fig. 10, a) and IgG-D "F00" antibody (fig. 10, B) compared to the chimeric variant canine IgG-a without Phe mutation (fig. 10, C) and the IgG-D without Phe mutation (fig. 10, D). Chimeric variant canine IgG-a and IgG-D antibodies with Phe mutations (fig. 10, a and B) showed enhanced association with canine FcRn at low pH (pH 6.0) and rapid dissociation at neutral pH (PBS pH 7.2). A similar enhanced binding profile was also observed in the case of the chimeric variant canine IgG-B "F00" antibody.
Example 16
Pharmacokinetics of Phe mutations in canine IgG
Pharmacokinetic analysis was performed using Spela-dawn rats. Rats were subcutaneously administered 2mg/kg of chimeric variant canine IgG-A "F00" antibody and chimeric variant canine IgG-A without Phe mutation (two rats per group). Serum samples were collected from rats before injection and at 0.5, 1, 6, 24,48, 72, 168, 216 and 336 hours after injection. The concentration of the canine chimeric antibody in the serum sample was determined by ELISA as follows.
The capture antibody (1. mu.g/mL in PBS) was coated on 96-well Maxisorp plates with 100. mu.l per well. Plates were incubated overnight at 4 ℃ and washed five times with PBST (PBS containing 0.05% Tween-20). Each well was blocked with 200 μ l 5% BSA in PBST and the plates were incubated at room temperature for 1 hour. Plates were washed five times with PBST. Dilutions of control antibody (1,000ng/mL to 0.1ng/mL) were added to the plates in duplicate and used with blank wells without control antibody to generate a standard curve. Serum samples were prepared at 10-, 20-and 40-fold dilutions in 5% BSA-PBST and added to the plates. Plates were incubated for 1 hour at room temperature and washed 5 times with PBST. Mu.l of HRP-conjugated antibody in 5% BSA-PBST (Bio-Rad, Cat. HCA204P) was added to each well at 0.25. mu.g/mL. Plates were incubated for 1 hour at room temperature and washed 5 times with PBST. Mu.l of QuantaBlu (Thermo Scientific, cat # 15169) was added to each well. After incubation for 10-15 minutes, fluorescence was measured at 325nm/420nm (emission/excitation). The titer against TNF α in the serum samples was calculated according to the standard curve.
AUC of IgG-A0-336hWas 150970, whereas IgG-a "F00" was 848924ng/mL _ h (fig. 11). The terminal half-life was estimated to be 33 hours and 152 hours, respectively. Thus, a single Phe mutation significantly improves the pharmacokinetic profile of the antibody in rats.
Example 17
Phe mutations in canine, feline, and equine IgG Fc
Interactions between Phe mutations and FcRn in canine IgG-A, IgG-B, IgG-C and IgG-D Fc were modeled using three-dimensional protein structure analysis. The aromatic side chain of Phe appears to have a hydrophobic interaction with canine FcRn at the Pro hydrophobic loop (pi-CH) of the "WPE" motif. In addition, the Phe hydrophobic side chain may be in direct contact with the Glu side chain alongside Pro of the same "WPE" motif. This interaction may result in energy loss if the Glu side chain deprotonates to a negative charge (e.g. at neutral pH). Thus, some level of protonation of the Glu residue may be required to minimize aromatic interaction with Glu-H. This may explain why the interaction between variant IgG with Phe mutation and FcRn decreases at neutral pH. Based on protein structure analysis, the interaction appeared to be conserved between the canine IgG-A, IgG-B, IgG-C and IgG-D Fc.
Furthermore, the interaction between Phe mutations in cat class IgG1a and IgG2Fc and cat class FcRn at complexation was modeled. The same interactions observed in the case of canine IgG Fc appear to be conserved for feline IgG Fc.
Interactions between Phe mutations in equine IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, and IgG7Fc and equine FcRn at the time of complexation were also modeled. Equine IgG Fc appears to maintain the same interaction.
Example 18
Other exemplary variants canine IgG Fc enhance canine FcRn interaction
The affinity of additional variant canine Fc polypeptides for FcRn was evaluated in the context of chimeric antibodies. Antibody variable light chains fused to canine kappa light chains and variable heavy chain sequences fused to wild-type IgG-B Fc polypeptide (comprising SEQ ID NO:35), variant canine IgG-B Fc polypeptide 0Y0 (comprising SEQ ID NO:203), variant canine IgG-B Fc polypeptide 0YH (comprising SEQ ID NO:204), variant canine IgG-B Fc polypeptide 0YY (comprising SEQ ID NO:205), and variant canine IgG-B Fc polypeptide 00Y (comprising SEQ ID NO:206) were expressed.
Binding analysis was performed using the biosensor OctetRed as follows. Briefly, biotinylated targets are captured on streptavidin sensor tips. The association of 20. mu.g/mL of antibody was with biotinylated target. The complexes were then used to bind canine FcRn (50 μ g/mL) at pH 6.0. Dissociation was performed at pH 7.2.
Each chimeric variant canine IgG-B antibody exhibited enhanced binding to canine FcRn at pH6.0, compared to the chimeric wild-type canine IgG-B antibody, and each had a considerable off-rate at neutral pH (fig. 12).
Example 19
Variant canine IgG Fc extends half-life of antibodies in vivo in canines
The in vivo half-life of variant canine Fc polypeptides against FcRn was evaluated in the context of chimeric antibodies. Antibody variable light chains fused to canine kappa light chains and variable heavy chains fused to wild-type IgG-B Fc polypeptide (comprising SEQ ID NO:35), variant canine IgG-B Fc polypeptide YTE (comprising SEQ ID NO:207), variant canine IgG-B Fc polypeptide 0Y0 (comprising SEQ ID NO:203), variant canine IgG-B Fc polypeptide F00 (comprising SEQ ID NO:197), variant canine IgG-B Fc polypeptide 0YH (comprising SEQ ID NO:204), and variant canine IgG-B Fc polypeptide Y00 (comprising SEQ ID NO:198) were expressed and purified to 40mg/mL in PBS (pH 7.2).
Canine pharmacokinetics is performed at the Absorption Systems, california company. Male beagle dogs (about 8-14kg) were obtained from Marshall Bioresources, North Rose, N.Y. A total of 12 dogs were used for the study, with each group of n-2 dogs. Six antibodies were administered subcutaneously to dogs at 4 mg/Kg. Serum samples were collected before injection and at 6, 24,48, 72, 96, 120, 144, 168, 216, 264, 336, 504, and 672 hours after injection. Canine chimeric antibody concentrations were determined by ELISA as described. Converting Cp between 144 hours and 336 hours to Ln [ Cp ]]Then fitted to Ln [ Cp ]]t＝-k*t+Ln[Cp]144hLinear equations of the form. The terminal half-life was then calculated from the slope k as listed in table 18 below. The 0Y0, F00, 0YH and Y00 mutations in the canine IgG-B Fc greatly improved the antibody inHalf-life in dog.
Table 18: effect of variant Canine IgG Fc on half-life of antibodies in dogs
Dog
Half-life period (Tian)
WT 1
13
WT 2
13
YTE 1
*NR
YTE 2
15
0Y0 1
*NR
0Y0 2
28
F00 1
*NR
F00 2
23
0YH 1
22
0YH 2
23
Y00 1
33
Y00 2
39
NR: the results are unreliable.
Sequence listing
<110> Kindrad biosciences GmbH
<120> medical NGF antagonist
<130> 01157-0018-00PCT
<150> US 62/821,438
<151> 2019-03-20
<160> 246
<170> PatentIn 3.5 edition
<210> 1
<211> 796
<212> PRT
<213> dog family (Canis lupus family)
<220>
<221> features not yet classified
<223> canine TrkA
<400> 1
Met Leu Arg Gly Gly Arg Leu Gly Gln Arg Gly Gly His Gly Arg Ala
1 5 10 15
Ala Gly Pro Gly Ser Leu Leu Ala Trp Leu Val Leu Ala Ser Ala Gly
20 25 30
Ala Ala Pro Cys Pro Asp Val Cys Cys Pro His Gly Pro Ser Gly Leu
35 40 45
Arg Cys Thr Arg Ala Gly Ala Leu Gln Ser Leu His Arg Leu Pro Gly
50 55 60
Val Glu Asn Leu Thr Glu Leu Tyr Ile Asp Asn Gln Glu His Leu Gln
65 70 75 80
His Leu Asp Ala Val His Leu Lys Gly Leu Gly Met Leu Arg Asp Leu
85 90 95
Thr Ile Val Lys Ser Gly Leu Arg Ser Val Ala Pro Asp Ala Phe His
100 105 110
Phe Thr Pro Arg Leu Arg Arg Leu Asn Leu Ser Phe Asn Ala Leu Glu
115 120 125
Ser Leu Ser Trp Lys Thr Val Gln Gly Leu Pro Leu Gln Glu Leu Val
130 135 140
Leu Ser Gly Asn Pro Leu His Cys Ser Cys Ala Leu His Trp Leu Leu
145 150 155 160
Arg Trp Glu Glu Glu Gly Leu Gly Gly Val Arg Gly Gln Arg Leu Gln
165 170 175
Cys Pro Gly Gln Gly Pro Leu Ala Leu Leu Ser Asn Ala Ser Cys Gly
180 185 190
Val Pro Val Leu Lys Val Gln Met Pro Asn Ala Ser Val Glu Val Gly
195 200 205
Asp Asp Val Leu Leu Gln Cys Gln Val Glu Gly Arg Gly Leu Glu Arg
210 215 220
Ala Gly Trp Ile Leu Pro Glu Val Glu Glu Leu Ala Thr Val Thr Pro
225 230 235 240
Ser Gly Asp Leu Pro Ser Leu Gly Leu Ile Leu Ala Asn Val Thr Ser
245 250 255
Asp Leu Asn Arg Lys Asn Val Thr Cys Trp Ala Glu Asn Asp Val Gly
260 265 270
Arg Ala Glu Val Ser Val Gln Val Asn Val Ser Phe Pro Ala Ser Val
275 280 285
Gln Leu His Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser
290 295 300
Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser
305 310 315 320
Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val
325 330 335
Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr
340 345 350
His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly
355 360 365
Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu
370 375 380
Phe Asn Pro Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr
385 390 395 400
Asn Ser Thr Ser Gly Asp Pro Val Glu Lys Lys Gly Gln Thr Pro Phe
405 410 415
Gly Val Ser Val Ala Val Gly Leu Ala Val Phe Ala Cys Leu Phe Leu
420 425 430
Ser Thr Leu Phe Leu Ala Leu Asn Lys Cys Gly Arg Arg Asn Lys Phe
435 440 445
Gly Gly Asn Arg Ala Val Val Leu Ala Pro Glu Asp Gly Leu Ala Met
450 455 460
Ser Leu His Phe Met Thr Leu Gly Gly Ser Ser Leu Ser Pro Thr Glu
465 470 475 480
Gly Lys Gly Ser Gly Leu Gln Gly His Ile Ile Glu Asn Pro Gln Tyr
485 490 495
Phe Ser Asp Ala Cys Val His His Ile Lys Arg Gln Asp Ile Val Leu
500 505 510
Lys Trp Glu Leu Gly Glu Gly Ala Phe Gly Lys Val Phe Leu Ala Glu
515 520 525
Cys His Asn Leu Leu Pro Glu Gln Asp Lys Met Leu Val Ala Val Lys
530 535 540
Ala Leu Lys Glu Val Ser Glu Ser Ala Arg Gln Asp Phe Gln Arg Glu
545 550 555 560
Ala Gln Leu Leu Thr Met Leu Gln His Gln His Ile Val Arg Phe Phe
565 570 575
Gly Val Cys Thr Glu Gly Arg Pro Leu Leu Met Val Phe Glu Tyr Met
580 585 590
Arg His Gly Asp Leu Asn Arg Phe Leu Arg Ser His Gly Pro Asp Ala
595 600 605
Lys Leu Leu Ala Gly Gly Glu Asp Val Ala Pro Gly Pro Leu Gly Leu
610 615 620
Gly Gln Leu Leu Ala Val Ala Ser Gln Val Ala Ala Gly Met Val Tyr
625 630 635 640
Leu Ala Gly Leu His Phe Val His Arg Asp Leu Ala Thr Arg Asn Cys
645 650 655
Leu Val Gly Gln Gly Leu Val Val Lys Ile Gly Asp Phe Gly Met Ser
660 665 670
Arg Asp Ile Tyr Ser Thr Asp Tyr Tyr Arg Val Gly Gly Arg Thr Met
675 680 685
Leu Pro Ile Arg Trp Met Pro Pro Glu Ser Ile Leu Tyr Arg Lys Phe
690 695 700
Thr Thr Glu Ser Asp Val Trp Ser Phe Gly Val Val Leu Trp Glu Ile
705 710 715 720
Phe Thr Tyr Gly Lys Gln Pro Trp Tyr Gln Leu Ser Asn Thr Glu Ala
725 730 735
Ile Glu Cys Ile Thr Gln Gly Arg Glu Leu Glu Arg Pro Arg Ala Cys
740 745 750
Pro Pro Glu Val Tyr Ala Ile Met Arg Gly Cys Trp Gln Arg Glu Pro
755 760 765
Gln Gln Arg His Ser Ile Lys Asp Val His Ala Arg Leu Gln Ala Leu
770 775 780
Ala Gln Ala Pro Pro Val Tyr Leu Asp Val Leu Gly
785 790 795
<210> 2
<211> 417
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD (v1)
<400> 2
Met Leu Arg Gly Gly Arg Leu Gly Gln Arg Gly Gly His Gly Arg Ala
1 5 10 15
Ala Gly Pro Gly Ser Leu Leu Ala Trp Leu Val Leu Ala Ser Ala Gly
20 25 30
Ala Ala Pro Cys Pro Asp Val Cys Cys Pro His Gly Pro Ser Gly Leu
35 40 45
Arg Cys Thr Arg Ala Gly Ala Leu Gln Ser Leu His Arg Leu Pro Gly
50 55 60
Val Glu Asn Leu Thr Glu Leu Tyr Ile Asp Asn Gln Glu His Leu Gln
65 70 75 80
His Leu Asp Ala Val His Leu Lys Gly Leu Gly Met Leu Arg Asp Leu
85 90 95
Thr Ile Val Lys Ser Gly Leu Arg Ser Val Ala Pro Asp Ala Phe His
100 105 110
Phe Thr Pro Arg Leu Arg Arg Leu Asn Leu Ser Phe Asn Ala Leu Glu
115 120 125
Ser Leu Ser Trp Lys Thr Val Gln Gly Leu Pro Leu Gln Glu Leu Val
130 135 140
Leu Ser Gly Asn Pro Leu His Cys Ser Cys Ala Leu His Trp Leu Leu
145 150 155 160
Arg Trp Glu Glu Glu Gly Leu Gly Gly Val Arg Gly Gln Arg Leu Gln
165 170 175
Cys Pro Gly Gln Gly Pro Leu Ala Leu Leu Ser Asn Ala Ser Cys Gly
180 185 190
Val Pro Val Leu Lys Val Gln Met Pro Asn Ala Ser Val Glu Val Gly
195 200 205
Asp Asp Val Leu Leu Gln Cys Gln Val Glu Gly Arg Gly Leu Glu Arg
210 215 220
Ala Gly Trp Ile Leu Pro Glu Val Glu Glu Leu Ala Thr Val Thr Pro
225 230 235 240
Ser Gly Asp Leu Pro Ser Leu Gly Leu Ile Leu Ala Asn Val Thr Ser
245 250 255
Asp Leu Asn Arg Lys Asn Val Thr Cys Trp Ala Glu Asn Asp Val Gly
260 265 270
Arg Ala Glu Val Ser Val Gln Val Asn Val Ser Phe Pro Ala Ser Val
275 280 285
Gln Leu His Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser
290 295 300
Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser
305 310 315 320
Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val
325 330 335
Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr
340 345 350
His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly
355 360 365
Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu
370 375 380
Phe Asn Pro Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr
385 390 395 400
Asn Ser Thr Ser Gly Asp Pro Val Glu Lys Lys Gly Gln Thr Pro Phe
405 410 415
Gly
<210> 3
<211> 125
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD (v2)
<400> 3
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp
115 120 125
<210> 4
<211> 101
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD (v3)
<400> 4
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro
100
<210> 5
<211> 99
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD (v4)
<400> 5
Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro
<210> 6
<211> 795
<212> PRT
<213> domestic Cat (Felis cat)
<220>
<221> features not yet classified
<223> feline TrkA
<400> 6
Met Leu Arg Gly Gly Arg Gly Pro Arg Gly Gly His Gly Arg Ala Ala
1 5 10 15
Gly Pro Gly Ser Leu Leu Ala Trp Leu Met Leu Ala Ser Ala Gly Ala
20 25 30
Ala Pro Cys Ala Asp Val Cys Cys Pro His Gly Pro Ser Gly Leu Arg
35 40 45
Cys Thr Arg Ala Gly Ala Leu Glu Ser Leu Arg Arg Leu Pro Gly Ala
50 55 60
Glu Asn Leu Thr Glu Leu Tyr Ile Glu Asn Gln Glu Arg Leu Arg His
65 70 75 80
Leu Glu Pro Ser Asp Leu Arg Gly Leu Gly Glu Leu Arg Gly Leu Thr
85 90 95
Ile Val Lys Ser Gly Leu Arg Phe Val Ala Pro Asp Val Phe His Phe
100 105 110
Thr Pro Arg Leu Ser Arg Leu Asn Leu Ser Phe Asn Ala Leu Glu Ser
115 120 125
Leu Ser Trp Lys Thr Val Gln Gly Leu Ser Leu Gln Glu Leu Val Leu
130 135 140
Ser Gly Asn Pro Leu Arg Cys Ser Cys Ala Leu His Trp Leu Leu Arg
145 150 155 160
Trp Glu Glu Glu Gly Leu Gly Gly Val Arg Ala Gln Arg Leu Gln Cys
165 170 175
Pro Gly Gln Gly Pro Leu Ala Leu Leu Ser Asn Ala Ser Cys Gly Val
180 185 190
Pro Val Leu Lys Val Gln Thr Pro Asn Ala Ser Val Asn Val Gly Asp
195 200 205
Asp Val Leu Leu Gln Cys Gln Val Glu Gly Arg Gly Leu Glu Gln Ala
210 215 220
Gly Trp Ile Leu Thr Glu Leu Glu Glu Ser Ala Thr Val Thr Gln Ser
225 230 235 240
Gly Ala Leu Pro Ser Leu Gly Leu Thr Leu Ala Asn Val Thr Ser Asp
245 250 255
Leu Asn Arg Lys Asn Val Thr Cys Trp Ala Glu Asn Asp Val Gly Arg
260 265 270
Ala Glu Val Ser Val Gln Val Asn Val Ser Phe Pro Ala Ser Val Gln
275 280 285
Leu His Ala Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val
290 295 300
Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val
305 310 315 320
Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Ala Ala
325 330 335
Asn Glu Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His
340 345 350
Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg
355 360 365
Ala Ala Ala Ser Val Leu Ala Ala Phe Met Asp Asn Pro Phe Glu Phe
370 375 380
Asn Pro Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Ser Asn
385 390 395 400
Ser Thr Ser Gly Asp Pro Val Glu Lys Lys Asp Glu Thr Pro Phe Gly
405 410 415
Val Ser Val Ala Val Gly Leu Ala Val Phe Ala Cys Leu Leu Leu Ser
420 425 430
Ala Phe Phe Leu Val Leu Asn Lys Cys Gly Arg Arg Asn Lys Phe Gly
435 440 445
Ile Asn Arg Thr Ala Val Leu Ala Pro Glu Asp Gly Leu Ala Met Ser
450 455 460
Leu His Phe Met Thr Leu Gly Gly Ser Ser Leu Ser Pro Thr Glu Gly
465 470 475 480
Lys Gly Ser Gly Leu Gln Gly His Ile Ile Glu Asn Pro Gln Tyr Phe
485 490 495
Ser Asp Ala Cys Val His His Ile Lys Arg Arg Asp Ile Val Leu Lys
500 505 510
Trp Glu Leu Gly Glu Gly Ala Phe Gly Lys Val Phe Leu Ala Glu Cys
515 520 525
Tyr Asn Leu Leu Pro Glu Gln Asp Lys Met Leu Val Ala Val Lys Ala
530 535 540
Leu Lys Glu Val Ser Glu Ser Ala Arg Gln Asp Phe Gln Arg Glu Ala
545 550 555 560
Gln Leu Leu Thr Val Leu Gln His Gln His Ile Val Arg Phe Phe Gly
565 570 575
Val Cys Thr Glu Gly Arg Pro Leu Leu Met Val Phe Glu Tyr Met Arg
580 585 590
His Gly Asp Leu Asn Arg Phe Leu Arg Ser His Gly Pro Asp Ala Lys
595 600 605
Leu Leu Ala Gly Arg Glu Asp Val Ala Pro Gly Pro Leu Gly Leu Ser
610 615 620
Gln Leu Leu Ala Val Ala Ser Gln Val Ala Ala Gly Met Val Tyr Leu
625 630 635 640
Ala Gly Leu His Phe Val His Arg Asp Leu Ala Thr Arg Asn Cys Leu
645 650 655
Val Gly Gln Gly Leu Val Val Lys Ile Gly Asp Phe Gly Met Ser Arg
660 665 670
Asp Ile Tyr Ser Thr Asp Tyr Tyr Arg Val Gly Gly Arg Thr Met Leu
675 680 685
Pro Ile Arg Trp Met Pro Pro Glu Ser Ile Leu Tyr Arg Lys Phe Thr
690 695 700
Thr Glu Ser Asp Val Trp Ser Phe Gly Val Val Leu Trp Glu Ile Phe
705 710 715 720
Thr Tyr Gly Lys Gln Pro Trp Tyr Gln Leu Ser Asn Thr Glu Ala Ile
725 730 735
Glu Cys Ile Thr Gln Gly Arg Glu Leu Glu Arg Pro Arg Ala Cys Pro
740 745 750
Pro Glu Val Tyr Ala Ile Met Arg Gly Cys Trp Gln Arg Glu Pro Gln
755 760 765
Gln Arg His Ser Ile Lys Glu Val His Ala Arg Leu Gln Ala Leu Ala
770 775 780
Gln Ala Pro Pro Val Tyr Leu Asp Val Leu Gly
785 790 795
<210> 7
<211> 416
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Cat TrkA ECD (v1)
<400> 7
Met Leu Arg Gly Gly Arg Gly Pro Arg Gly Gly His Gly Arg Ala Ala
1 5 10 15
Gly Pro Gly Ser Leu Leu Ala Trp Leu Met Leu Ala Ser Ala Gly Ala
20 25 30
Ala Pro Cys Ala Asp Val Cys Cys Pro His Gly Pro Ser Gly Leu Arg
35 40 45
Cys Thr Arg Ala Gly Ala Leu Glu Ser Leu Arg Arg Leu Pro Gly Ala
50 55 60
Glu Asn Leu Thr Glu Leu Tyr Ile Glu Asn Gln Glu Arg Leu Arg His
65 70 75 80
Leu Glu Pro Ser Asp Leu Arg Gly Leu Gly Glu Leu Arg Gly Leu Thr
85 90 95
Ile Val Lys Ser Gly Leu Arg Phe Val Ala Pro Asp Val Phe His Phe
100 105 110
Thr Pro Arg Leu Ser Arg Leu Asn Leu Ser Phe Asn Ala Leu Glu Ser
115 120 125
Leu Ser Trp Lys Thr Val Gln Gly Leu Ser Leu Gln Glu Leu Val Leu
130 135 140
Ser Gly Asn Pro Leu Arg Cys Ser Cys Ala Leu His Trp Leu Leu Arg
145 150 155 160
Trp Glu Glu Glu Gly Leu Gly Gly Val Arg Ala Gln Arg Leu Gln Cys
165 170 175
Pro Gly Gln Gly Pro Leu Ala Leu Leu Ser Asn Ala Ser Cys Gly Val
180 185 190
Pro Val Leu Lys Val Gln Thr Pro Asn Ala Ser Val Asn Val Gly Asp
195 200 205
Asp Val Leu Leu Gln Cys Gln Val Glu Gly Arg Gly Leu Glu Gln Ala
210 215 220
Gly Trp Ile Leu Thr Glu Leu Glu Glu Ser Ala Thr Val Thr Gln Ser
225 230 235 240
Gly Ala Leu Pro Ser Leu Gly Leu Thr Leu Ala Asn Val Thr Ser Asp
245 250 255
Leu Asn Arg Lys Asn Val Thr Cys Trp Ala Glu Asn Asp Val Gly Arg
260 265 270
Ala Glu Val Ser Val Gln Val Asn Val Ser Phe Pro Ala Ser Val Gln
275 280 285
Leu His Ala Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val
290 295 300
Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val
305 310 315 320
Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Ala Ala
325 330 335
Asn Glu Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His
340 345 350
Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg
355 360 365
Ala Ala Ala Ser Val Leu Ala Ala Phe Met Asp Asn Pro Phe Glu Phe
370 375 380
Asn Pro Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Ser Asn
385 390 395 400
Ser Thr Ser Gly Asp Pro Val Glu Lys Lys Asp Glu Thr Pro Phe Gly
405 410 415
<210> 8
<211> 125
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Cat TrkA ECD (v2)
<400> 8
Val Ser Phe Pro Ala Ser Val Gln Leu His Ala Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Ala Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Ser Val Leu Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Ser Asn Ser Thr Ser Gly Asp
115 120 125
<210> 9
<211> 101
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Cat TrkA ECD (v3)
<400> 9
Val Ser Phe Pro Ala Ser Val Gln Leu His Ala Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Ala Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Ser Val Leu Ala Ala
85 90 95
Phe Met Asp Asn Pro
100
<210> 10
<211> 99
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Cat TrkA ECD (v4)
<400> 10
Phe Pro Ala Ser Val Gln Leu His Ala Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Ala Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Ser Val Leu Ala Ala Phe Met
85 90 95
Asp Asn Pro
<210> 11
<211> 796
<212> PRT
<213> horse (Equus caballus)
<220>
<221> features not yet classified
<223> equine TrkA
<400> 11
Met Leu Arg Gly Gly Arg Arg Gly Gln Leu Gly Trp His Gly Arg Ala
1 5 10 15
Thr Gly Pro Gly Ser Leu Leu Ala Trp Leu Met Leu Ala Ser Ala Gly
20 25 30
Ala Ala Pro Cys Pro Asp Ala Cys Cys Pro His Gly Pro Ser Gly Leu
35 40 45
Arg Cys Thr Gln Pro Gly Ala Leu Asp Ser Leu Arg His Leu Pro Gly
50 55 60
Ala Glu Asn Leu Thr Glu Leu Tyr Ile Glu Asn Gln Gln Asn Leu Gln
65 70 75 80
Arg Leu Glu Arg Asn Asp Leu Arg Gly Leu Gly Glu Leu Arg Asn Leu
85 90 95
Thr Ile Val Lys Ser Gly Leu His Phe Val Ala Pro Asp Ala Phe His
100 105 110
Phe Thr Pro Arg Leu Ser Arg Leu Asn Leu Ser Phe Asn Ala Leu Lys
115 120 125
Ser Leu Ser Trp Lys Thr Val Gln Gly Leu Ser Leu Gln Gln Leu Val
130 135 140
Leu Leu Gly Asn Pro Leu Asp Cys Ser Cys Ala Leu Arg Trp Leu Gln
145 150 155 160
Arg Trp Glu Glu Glu Gly Leu Gly Gly Val Arg Glu Gln Lys Leu Gln
165 170 175
Cys His Gln Gln Gly Pro Leu Ala Leu Met Ser Asn Thr Asn Cys Gly
180 185 190
Val Pro Leu Leu Lys Val Gln Val Pro Asn Ala Ser Val Asp Val Gly
195 200 205
Asp Asn Val Trp Leu Gln Cys Gln Val Glu Gly Gln Gly Leu Glu Gln
210 215 220
Ala Gly Trp Ile Leu Thr Glu Leu Glu Glu Ser Ala Thr Val Met Gln
225 230 235 240
Ser Gly Gly Leu Pro Ser Leu Gly Leu Thr Leu Ala Asn Val Thr Ser
245 250 255
Asp Leu Asn Arg Lys Asn Val Thr Cys Trp Ala Glu Asn Asp Val Gly
260 265 270
Arg Ala Glu Val Ser Val Gln Val Asn Val Ser Phe Pro Ala Ser Val
275 280 285
His Leu Gln Thr Ala Val Glu Gln His His Trp Cys Ile Pro Phe Ser
290 295 300
Val Asp Gly Gln Pro Ala Pro Thr Leu Arg Trp Leu Phe Asn Gly Ser
305 310 315 320
Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Ser Ala
325 330 335
Ala Asn Glu Thr Met Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr
340 345 350
His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala Thr Asn Pro Tyr Gly
355 360 365
Gln Asp Ser Ala Ser Val Met Val Ala Phe Met Asp Asn Pro Phe Glu
370 375 380
Phe Asn Pro Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr
385 390 395 400
Asn Ser Thr Ser Arg Asp Pro Val Glu Lys Lys Asp Glu Thr His Phe
405 410 415
Gly Val Ser Val Ala Val Gly Leu Ala Val Phe Ala Cys Leu Phe Leu
420 425 430
Ser Thr Leu Phe Leu Val Leu Asn Lys Cys Gly Arg Arg Asn Lys Phe
435 440 445
Gly Ile Asn Arg Pro Ala Val Leu Ala Pro Glu Asp Gly Leu Ala Met
450 455 460
Ser Leu His Phe Met Thr Leu Gly Gly Ser Ser Leu Ser Pro Thr Glu
465 470 475 480
Gly Lys Gly Ser Gly Leu Gln Gly His Ile Ile Glu Asn Pro Gln Tyr
485 490 495
Phe Ser Asp Ala Cys Val His His Ile Lys Arg Arg Asp Ile Val Leu
500 505 510
Lys Trp Glu Leu Gly Glu Gly Ala Phe Gly Lys Val Phe Leu Ala Glu
515 520 525
Cys His Asn Leu Leu Pro Glu Gln Asp Lys Met Leu Val Ala Val Lys
530 535 540
Ala Leu Lys Glu Val Ser Glu Ser Ala Arg Gln Asp Phe Gln Arg Glu
545 550 555 560
Ala Glu Leu Leu Thr Met Leu Gln His Gln His Ile Val Arg Phe Phe
565 570 575
Gly Val Cys Thr Glu Gly Arg Pro Leu Leu Met Val Phe Glu Tyr Met
580 585 590
Arg His Gly Asp Leu Asn Arg Phe Leu Arg Ser His Gly Pro Asp Ala
595 600 605
Lys Leu Leu Ala Gly Gly Glu Asp Val Ala Pro Gly Pro Leu Gly Leu
610 615 620
Gly Gln Leu Leu Ala Val Ala Ser Gln Val Ala Ala Gly Met Val Tyr
625 630 635 640
Leu Ala Gly Leu His Phe Val His Arg Asp Leu Ala Thr Arg Asn Cys
645 650 655
Leu Val Gly Gln Gly Leu Val Val Lys Ile Gly Asp Phe Gly Met Ser
660 665 670
Arg Asp Ile Tyr Ser Thr Asp Tyr Tyr Arg Val Gly Gly Arg Thr Met
675 680 685
Leu Pro Ile Arg Trp Met Pro Pro Glu Ser Ile Leu Tyr Arg Lys Phe
690 695 700
Thr Thr Glu Ser Asp Val Trp Ser Phe Gly Val Val Leu Trp Glu Ile
705 710 715 720
Phe Thr Tyr Gly Lys Gln Pro Trp Tyr Gln Leu Ser Asn Thr Glu Ala
725 730 735
Ile Glu Cys Ile Thr Gln Gly Arg Glu Leu Glu Arg Pro Arg Ala Cys
740 745 750
Pro Pro Glu Val Tyr Ala Ile Met Arg Gly Cys Trp Gln Arg Glu Pro
755 760 765
Gln Gln Arg His Ser Ile Lys Asp Val His Ala Arg Leu Gln Ala Leu
770 775 780
Val Gln Ala Pro Pro Val Tyr Leu Asp Val Leu Gly
785 790 795
<210> 12
<211> 417
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD (v1)
<400> 12
Met Leu Arg Gly Gly Arg Arg Gly Gln Leu Gly Trp His Gly Arg Ala
1 5 10 15
Thr Gly Pro Gly Ser Leu Leu Ala Trp Leu Met Leu Ala Ser Ala Gly
20 25 30
Ala Ala Pro Cys Pro Asp Ala Cys Cys Pro His Gly Pro Ser Gly Leu
35 40 45
Arg Cys Thr Gln Pro Gly Ala Leu Asp Ser Leu Arg His Leu Pro Gly
50 55 60
Ala Glu Asn Leu Thr Glu Leu Tyr Ile Glu Asn Gln Gln Asn Leu Gln
65 70 75 80
Arg Leu Glu Arg Asn Asp Leu Arg Gly Leu Gly Glu Leu Arg Asn Leu
85 90 95
Thr Ile Val Lys Ser Gly Leu His Phe Val Ala Pro Asp Ala Phe His
100 105 110
Phe Thr Pro Arg Leu Ser Arg Leu Asn Leu Ser Phe Asn Ala Leu Lys
115 120 125
Ser Leu Ser Trp Lys Thr Val Gln Gly Leu Ser Leu Gln Gln Leu Val
130 135 140
Leu Leu Gly Asn Pro Leu Asp Cys Ser Cys Ala Leu Arg Trp Leu Gln
145 150 155 160
Arg Trp Glu Glu Glu Gly Leu Gly Gly Val Arg Glu Gln Lys Leu Gln
165 170 175
Cys His Gln Gln Gly Pro Leu Ala Leu Met Ser Asn Thr Asn Cys Gly
180 185 190
Val Pro Leu Leu Lys Val Gln Val Pro Asn Ala Ser Val Asp Val Gly
195 200 205
Asp Asn Val Trp Leu Gln Cys Gln Val Glu Gly Gln Gly Leu Glu Gln
210 215 220
Ala Gly Trp Ile Leu Thr Glu Leu Glu Glu Ser Ala Thr Val Met Gln
225 230 235 240
Ser Gly Gly Leu Pro Ser Leu Gly Leu Thr Leu Ala Asn Val Thr Ser
245 250 255
Asp Leu Asn Arg Lys Asn Val Thr Cys Trp Ala Glu Asn Asp Val Gly
260 265 270
Arg Ala Glu Val Ser Val Gln Val Asn Val Ser Phe Pro Ala Ser Val
275 280 285
His Leu Gln Thr Ala Val Glu Gln His His Trp Cys Ile Pro Phe Ser
290 295 300
Val Asp Gly Gln Pro Ala Pro Thr Leu Arg Trp Leu Phe Asn Gly Ser
305 310 315 320
Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Ser Ala
325 330 335
Ala Asn Glu Thr Met Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr
340 345 350
His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala Thr Asn Pro Tyr Gly
355 360 365
Gln Asp Ser Ala Ser Val Met Val Ala Phe Met Asp Asn Pro Phe Glu
370 375 380
Phe Asn Pro Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr
385 390 395 400
Asn Ser Thr Ser Arg Asp Pro Val Glu Lys Lys Asp Glu Thr His Phe
405 410 415
Gly
<210> 13
<211> 125
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD (v2)
<400> 13
Val Ser Phe Pro Ala Ser Val His Leu Gln Thr Ala Val Glu Gln His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Thr Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Ser Ala Ala Asn Glu Thr Met Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser Val Met Val Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Arg Asp
115 120 125
<210> 14
<211> 101
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD (v3)
<400> 14
Val Ser Phe Pro Ala Ser Val His Leu Gln Thr Ala Val Glu Gln His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Thr Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Ser Ala Ala Asn Glu Thr Met Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser Val Met Val Ala
85 90 95
Phe Met Asp Asn Pro
100
<210> 15
<211> 99
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD (v4)
<400> 15
Phe Pro Ala Ser Val His Leu Gln Thr Ala Val Glu Gln His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Thr Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Ser Ala Ala Asn Glu Thr Met Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser Val Met Val Ala Phe Met
85 90 95
Asp Asn Pro
<210> 16
<211> 384
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-wild type canine with signal sequence
IgG-B Fc protein A + C1q + CD16+
<400> 16
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro
115 120 125
Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr
130 135 140
Ser Gly Asp Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
145 150 155 160
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
165 170 175
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
180 185 190
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
195 200 205
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
210 215 220
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
225 230 235 240
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
245 250 255
Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
260 265 270
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
275 280 285
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
290 295 300
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
305 310 315 320
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
325 330 335
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
340 345 350
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
355 360 365
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
370 375 380
<210> 17
<211> 360
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-wild type canine with signal sequence
IgG-B Fc protein A + C1q + CD16+
<400> 17
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu
130 135 140
Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
145 150 155 160
Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
165 170 175
Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly
180 185 190
Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn
195 200 205
Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp
210 215 220
Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala Leu Pro
225 230 235 240
Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln
245 250 255
Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn
260 265 270
Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile
275 280 285
Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr
290 295 300
Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr
305 310 315 320
Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe
325 330 335
Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu
340 345 350
Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 18
<211> 358
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-wild type canine with signal sequence
IgG-B Fc protein A + C1q + CD16+
<400> 18
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Phe Pro Ala Ser Val Gln Leu His Glu Ala
20 25 30
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe
100 105 110
Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
115 120 125
Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu
130 135 140
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu
145 150 155 160
Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp
165 170 175
Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln
180 185 190
Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr
195 200 205
Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys
210 215 220
Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala Leu Pro Ser Pro
225 230 235 240
Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser
245 250 255
Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val
260 265 270
Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val
275 280 285
Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr
290 295 300
Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
305 310 315 320
Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys
325 330 335
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu
340 345 350
Ser His Ser Pro Gly Lys
355
<210> 19
<211> 393
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Cat TrkA ECD v 2-wild type Cat with Signal sequence
IgG2Fc protein A + C1q-CD16-
<400> 19
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Ala Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Ala Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Ser Val Leu Ala Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro
115 120 125
Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Ser Asn Ser Thr
130 135 140
Ser Gly Asp Ser Gly Gly Gly Ser Gly Gly Gly Ser Pro Lys Thr Ala
145 150 155 160
Ser Thr Ile Glu Ser Lys Thr Gly Glu Gly Pro Lys Cys Pro Val Pro
165 170 175
Glu Ile Pro Gly Ala Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys
180 185 190
Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr Cys Leu Val Val
195 200 205
Asp Leu Gly Pro Asp Asp Ser Asn Val Gln Ile Thr Trp Phe Val Asp
210 215 220
Asn Thr Glu Met His Thr Ala Lys Thr Arg Pro Arg Glu Glu Gln Phe
225 230 235 240
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile Leu His Gln Asp
245 250 255
Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn Ser Lys Ser Leu
260 265 270
Pro Ser Ala Met Glu Arg Thr Ile Ser Lys Ala Lys Gly Gln Pro His
275 280 285
Glu Pro Gln Val Tyr Val Leu Pro Pro Thr Gln Glu Glu Leu Ser Glu
290 295 300
Asn Lys Val Ser Val Thr Cys Leu Ile Lys Gly Phe His Pro Pro Asp
305 310 315 320
Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu Pro Glu Asn Asn
325 330 335
Tyr Gln Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly Thr Tyr Phe Leu
340 345 350
Tyr Ser Arg Leu Ser Val Asp Arg Ser His Trp Gln Arg Gly Asn Thr
355 360 365
Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser His His Thr Gln
370 375 380
Lys Ser Leu Thr Gln Ser Pro Gly Lys
385 390
<210> 20
<211> 369
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Cat TrkA ECD v 3-wild type Cat with Signal sequence
IgG2Fc protein A + C1q-CD16-
<400> 20
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Ala Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Ala Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Ser Val Leu Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Pro Lys Thr Ala Ser Thr Ile Glu Ser Lys Thr Gly
130 135 140
Glu Gly Pro Lys Cys Pro Val Pro Glu Ile Pro Gly Ala Pro Ser Val
145 150 155 160
Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr
165 170 175
Pro Glu Val Thr Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asn
180 185 190
Val Gln Ile Thr Trp Phe Val Asp Asn Thr Glu Met His Thr Ala Lys
195 200 205
Thr Arg Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser
210 215 220
Val Leu Pro Ile Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys
225 230 235 240
Cys Lys Val Asn Ser Lys Ser Leu Pro Ser Ala Met Glu Arg Thr Ile
245 250 255
Ser Lys Ala Lys Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro
260 265 270
Pro Thr Gln Glu Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu
275 280 285
Ile Lys Gly Phe His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr
290 295 300
Gly Gln Pro Glu Pro Glu Asn Asn Tyr Gln Thr Thr Pro Pro Gln Leu
305 310 315 320
Asp Ser Asp Gly Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg
325 330 335
Ser His Trp Gln Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu
340 345 350
Ala Leu His Ser His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly
355 360 365
Lys
<210> 21
<211> 367
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Cat TrkA ECD v 4-wild type Cat with Signal sequence
IgG2Fc protein A + C1q-CD16-
<400> 21
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Phe Pro Ala Ser Val Gln Leu His Ala Ala
20 25 30
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Ala Ala Asn Glu Thr Val
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Ser
100 105 110
Val Leu Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
115 120 125
Gly Ser Pro Lys Thr Ala Ser Thr Ile Glu Ser Lys Thr Gly Glu Gly
130 135 140
Pro Lys Cys Pro Val Pro Glu Ile Pro Gly Ala Pro Ser Val Phe Ile
145 150 155 160
Phe Pro Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu
165 170 175
Val Thr Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asn Val Gln
180 185 190
Ile Thr Trp Phe Val Asp Asn Thr Glu Met His Thr Ala Lys Thr Arg
195 200 205
Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu
210 215 220
Pro Ile Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys
225 230 235 240
Val Asn Ser Lys Ser Leu Pro Ser Ala Met Glu Arg Thr Ile Ser Lys
245 250 255
Ala Lys Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Thr
260 265 270
Gln Glu Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys
275 280 285
Gly Phe His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln
290 295 300
Pro Glu Pro Glu Asn Asn Tyr Gln Thr Thr Pro Pro Gln Leu Asp Ser
305 310 315 320
Asp Gly Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser His
325 330 335
Trp Gln Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu
340 345 350
His Ser His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
355 360 365
<210> 22
<211> 389
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD v 2-variant equine with Signal sequence
IgG2Fc protein A + C1q-CD16-
<400> 22
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Val Ser Phe Pro Ala Ser Val His Leu Gln Thr Ala
20 25 30
Val Glu Gln His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Thr Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Ser Ala Ala Asn Glu Thr Met
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser
100 105 110
Val Met Val Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp
115 120 125
Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Arg
130 135 140
Asp Pro Pro Cys Val Leu Ser Ala Glu Gly Val Ile Pro Ile Pro Ser
145 150 155 160
Val Pro Lys Pro Gln Cys Pro Pro Tyr Thr His Ser Lys Phe Leu Gly
165 170 175
Gly Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu Met
180 185 190
Ile Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp
195 200 205
Gln Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val
210 215 220
His Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr
225 230 235 240
Arg Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly
245 250 255
Lys Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile
260 265 270
Ser Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val
275 280 285
Tyr Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser
290 295 300
Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu
305 310 315 320
Trp Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr
325 330 335
Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu
340 345 350
Ser Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala
355 360 365
Val Met His Glu Ala Leu His Asn His Tyr Thr Lys Thr Asp Ile Ser
370 375 380
Glu Ser Leu Gly Lys
385
<210> 23
<211> 365
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD v 3-variant equine with Signal sequence
IgG2Fc protein A + C1q-CD16-
<400> 23
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Val Ser Phe Pro Ala Ser Val His Leu Gln Thr Ala
20 25 30
Val Glu Gln His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Thr Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Ser Ala Ala Asn Glu Thr Met
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser
100 105 110
Val Met Val Ala Phe Met Asp Asn Pro Pro Pro Cys Val Leu Ser Ala
115 120 125
Glu Gly Val Ile Pro Ile Pro Ser Val Pro Lys Pro Gln Cys Pro Pro
130 135 140
Tyr Thr His Ser Lys Phe Leu Gly Gly Pro Ser Val Phe Ile Phe Pro
145 150 155 160
Pro Asn Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Val Val Thr
165 170 175
Cys Val Val Val Asn Leu Ser Asp Gln Tyr Pro Asp Val Gln Phe Ser
180 185 190
Trp Tyr Val Asp Asn Thr Glu Val His Ser Ala Ile Thr Lys Gln Arg
195 200 205
Glu Ala Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
210 215 220
Gln His Gln Asp Trp Leu Ser Gly Lys Glu Phe Lys Cys Ser Val Thr
225 230 235 240
Asn Val Gly Val Pro Gln Pro Ile Ser Arg Ala Ile Ser Arg Gly Lys
245 250 255
Gly Pro Ser Arg Val Pro Gln Val Tyr Val Leu Pro Pro His Pro Asp
260 265 270
Glu Leu Ala Lys Ser Lys Val Ser Val Thr Cys Leu Val Lys Asp Phe
275 280 285
Tyr Pro Pro Asp Ile Ser Val Glu Trp Gln Ser Asn Arg Trp Pro Glu
290 295 300
Leu Glu Gly Lys Tyr Ser Thr Thr Pro Ala Gln Leu Asp Gly Asp Gly
305 310 315 320
Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Leu Glu Thr Ser Arg Trp Gln
325 330 335
Gln Val Glu Ser Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn
340 345 350
His Tyr Thr Lys Thr Asp Ile Ser Glu Ser Leu Gly Lys
355 360 365
<210> 24
<211> 363
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD v 4-variant equine with Signal sequence
IgG2Fc protein A + C1q-CD16-
<400> 24
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Phe Pro Ala Ser Val His Leu Gln Thr Ala Val Glu
20 25 30
Gln His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro
35 40 45
Thr Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe
50 55 60
Ile Phe Thr Glu Phe Leu Glu Ser Ala Ala Asn Glu Thr Met Arg His
65 70 75 80
Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr
85 90 95
Thr Leu Leu Ala Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser Val Met
100 105 110
Val Ala Phe Met Asp Asn Pro Pro Pro Cys Val Leu Ser Ala Glu Gly
115 120 125
Val Ile Pro Ile Pro Ser Val Pro Lys Pro Gln Cys Pro Pro Tyr Thr
130 135 140
His Ser Lys Phe Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Asn
145 150 155 160
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Val Val Thr Cys Val
165 170 175
Val Val Asn Leu Ser Asp Gln Tyr Pro Asp Val Gln Phe Ser Trp Tyr
180 185 190
Val Asp Asn Thr Glu Val His Ser Ala Ile Thr Lys Gln Arg Glu Ala
195 200 205
Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gln His
210 215 220
Gln Asp Trp Leu Ser Gly Lys Glu Phe Lys Cys Ser Val Thr Asn Val
225 230 235 240
Gly Val Pro Gln Pro Ile Ser Arg Ala Ile Ser Arg Gly Lys Gly Pro
245 250 255
Ser Arg Val Pro Gln Val Tyr Val Leu Pro Pro His Pro Asp Glu Leu
260 265 270
Ala Lys Ser Lys Val Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro
275 280 285
Pro Asp Ile Ser Val Glu Trp Gln Ser Asn Arg Trp Pro Glu Leu Glu
290 295 300
Gly Lys Tyr Ser Thr Thr Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr
305 310 315 320
Phe Leu Tyr Ser Lys Leu Ser Leu Glu Thr Ser Arg Trp Gln Gln Val
325 330 335
Glu Ser Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn His Tyr
340 345 350
Thr Lys Thr Asp Ile Ser Glu Ser Leu Gly Lys
355 360
<210> 25
<211> 125
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD (v2) additional disulfide
<400> 25
Val Ser Phe Pro Ala Ser Cys Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Cys Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp
115 120 125
<210> 26
<211> 101
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD (v3) additional disulfide
<400> 26
Val Ser Phe Pro Ala Ser Cys Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Cys Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro
100
<210> 27
<211> 99
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD (v4) additional disulfide
<400> 27
Phe Pro Ala Ser Cys Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Cys Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro
<210> 28
<211> 125
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Cat TrkA ECD (v2) additional disulfides
<400> 28
Val Ser Phe Pro Ala Ser Cys Gln Leu His Ala Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Ala Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Cys Ala Ala Ser Val Leu Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Ser Asn Ser Thr Ser Gly Asp
115 120 125
<210> 29
<211> 101
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Cat TrkA ECD (v3) additional disulfides
<400> 29
Val Ser Phe Pro Ala Ser Cys Gln Leu His Ala Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Ala Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Cys Ala Ala Ser Val Leu Ala Ala
85 90 95
Phe Met Asp Asn Pro
100
<210> 30
<211> 99
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Cat TrkA ECD (v4) additional disulfides
<400> 30
Phe Pro Ala Ser Cys Gln Leu His Ala Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Ala Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Cys Ala Ala Ser Val Leu Ala Ala Phe Met
85 90 95
Asp Asn Pro
<210> 31
<211> 125
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD (v2) additional disulfides
<400> 31
Val Ser Phe Pro Ala Ser Cys His Leu Gln Thr Ala Val Glu Gln His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Thr Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Ser Ala Ala Asn Glu Thr Met Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Thr Asn Pro Tyr Gly Gln Cys Ser Ala Ser Val Met Val Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Arg Asp
115 120 125
<210> 32
<211> 101
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD (v3) additional disulfides
<400> 32
Val Ser Phe Pro Ala Ser Cys His Leu Gln Thr Ala Val Glu Gln His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Thr Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Ser Ala Ala Asn Glu Thr Met Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Thr Asn Pro Tyr Gly Gln Cys Ser Ala Ser Val Met Val Ala
85 90 95
Phe Met Asp Asn Pro
100
<210> 33
<211> 99
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD (v4) additional disulfides
<400> 33
Phe Pro Ala Ser Cys His Leu Gln Thr Ala Val Glu Gln His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Thr Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Ser Ala Ala Asn Glu Thr Met Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Thr Asn Pro Tyr Gly Gln Cys Ser Ala Ser Val Met Val Ala Phe Met
85 90 95
Asp Asn Pro
<210> 34
<211> 221
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type Canine IgG-A Fc protein A-C1q
- CD16 -
<400> 34
Pro Val Pro Glu Pro Leu Gly Gly Pro Ser Val Leu Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Ile Leu Arg Ile Thr Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Leu Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Ser Arg Glu
50 55 60
Gln Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Glu
65 70 75 80
His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val Asn His
85 90 95
Ile Asp Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Arg Ala His Lys Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys Glu
115 120 125
Leu Ser Ser Ser Asp Thr Val Ser Ile Thr Cys Leu Ile Lys Asp Phe
130 135 140
Tyr Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu
145 150 155 160
Pro Glu Arg Lys His Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly
165 170 175
Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln
180 185 190
Gln Gly Asp Pro Phe Thr Cys Ala Val Met His Glu Thr Leu Gln Asn
195 200 205
His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 35
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type Canine IgG-B Fc protein A + C1q
+ CD16 +
<400> 35
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 36
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type canine IgG-B Fc with hinge
Protein A + C1q + CD16+
<400> 36
Pro Lys Arg Glu Asn Gly Arg Val Pro Arg Pro Pro Asp Cys Pro Lys
1 5 10 15
Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr
35 40 45
Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser
50 55 60
Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn
100 105 110
Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg
115 120 125
Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu
130 135 140
Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe
145 150 155 160
Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu
165 170 175
Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly
180 185 190
Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln
195 200 205
Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn
210 215 220
His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
225 230 235
<210> 37
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type Canine IgG-C Fc protein A-C1q
+ CD16 +
<400> 37
Pro Gly Cys Gly Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Ile Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asn Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu
50 55 60
Glu Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly
100 105 110
Gln Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu
115 120 125
Met Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe
130 135 140
Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 38
<211> 235
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type canine IgG-C Fc with hinge
Protein A-C1q + CD16+
<400> 38
Ala Lys Glu Cys Glu Cys Lys Cys Asn Cys Asn Asn Cys Pro Cys Pro
1 5 10 15
Gly Cys Gly Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys
20 25 30
Pro Lys Asp Ile Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys Val
35 40 45
Val Val Asp Leu Asp Pro Glu Asn Pro Glu Val Gln Ile Ser Trp Phe
50 55 60
Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu Glu
65 70 75 80
Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His
85 90 95
Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Lys Val Asn Asn Lys
100 105 110
Ala Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly Gln
115 120 125
Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu Met
130 135 140
Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe Pro
145 150 155 160
Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu
165 170 175
Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr
180 185 190
Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly
195 200 205
Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr
210 215 220
Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
225 230 235
<210> 39
<211> 221
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type Canine IgG-D Fc protein A-C1q
- CD16 -
<400> 39
Pro Val Pro Glu Ser Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Ile Leu Arg Ile Thr Arg Thr Pro Glu Ile Thr Cys
20 25 30
Val Val Leu Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Gln Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile Glu
65 70 75 80
His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val Asn His
85 90 95
Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys Glu
115 120 125
Leu Ser Ser Ser Asp Thr Val Thr Leu Thr Cys Leu Ile Lys Asp Phe
130 135 140
Phe Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Pro Glu
145 150 155 160
Pro Glu Ser Lys Tyr His Thr Thr Ala Pro Gln Leu Asp Glu Asp Gly
165 170 175
Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln
180 185 190
Gln Gly Asp Thr Phe Thr Cys Ala Val Met His Glu Ala Leu Gln Asn
195 200 205
His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 40
<211> 221
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-A Fc C1q-CD16-
Protein A + I (21) T R (23) L T (25) A E (80) G T (205) A Q (207) H
<400> 40
Pro Val Pro Glu Pro Leu Gly Gly Pro Ser Val Leu Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Leu Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Ser Arg Glu
50 55 60
Gln Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val Asn His
85 90 95
Ile Asp Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Arg Ala His Lys Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys Glu
115 120 125
Leu Ser Ser Ser Asp Thr Val Ser Ile Thr Cys Leu Ile Lys Asp Phe
130 135 140
Tyr Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu
145 150 155 160
Pro Glu Arg Lys His Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly
165 170 175
Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln
180 185 190
Gln Gly Asp Pro Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn
195 200 205
His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 41
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-C Fc C1q + protein A +
I(21)T V(23)L T(24)I
<400> 41
Pro Gly Cys Gly Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Thr Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asn Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu
50 55 60
Glu Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly
100 105 110
Gln Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu
115 120 125
Met Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe
130 135 140
Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 42
<211> 221
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-D Fc C1q-CD16-
Protein A + I (21) T R (23) L T (25) A E (80) G Q (207) H
<400> 42
Pro Val Pro Glu Ser Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Ile Thr Cys
20 25 30
Val Val Leu Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Gln Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val Asn His
85 90 95
Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys Glu
115 120 125
Leu Ser Ser Ser Asp Thr Val Thr Leu Thr Cys Leu Ile Lys Asp Phe
130 135 140
Phe Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Pro Glu
145 150 155 160
Pro Glu Ser Lys Tyr His Thr Thr Ala Pro Gln Leu Asp Glu Asp Gly
165 170 175
Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln
180 185 190
Gln Gly Asp Thr Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn
195 200 205
His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 43
<211> 221
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-A Fc C1q-CD16-
Protein A + I (21) T Q (207) H
<400> 43
Pro Val Pro Glu Pro Leu Gly Gly Pro Ser Val Leu Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Arg Ile Thr Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Leu Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Ser Arg Glu
50 55 60
Gln Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Glu
65 70 75 80
His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val Asn His
85 90 95
Ile Asp Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Arg Ala His Lys Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys Glu
115 120 125
Leu Ser Ser Ser Asp Thr Val Ser Ile Thr Cys Leu Ile Lys Asp Phe
130 135 140
Tyr Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu
145 150 155 160
Pro Glu Arg Lys His Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly
165 170 175
Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln
180 185 190
Gln Gly Asp Pro Phe Thr Cys Ala Val Met His Glu Thr Leu His Asn
195 200 205
His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 44
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-C Fc C1q + protein A +
I(21)T
<400> 44
Pro Gly Cys Gly Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asn Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu
50 55 60
Glu Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly
100 105 110
Gln Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu
115 120 125
Met Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe
130 135 140
Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 45
<211> 221
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-D Fc C1 q-protein A +
I(21)T Q(207)H
<400> 45
Pro Val Pro Glu Ser Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Arg Ile Thr Arg Thr Pro Glu Ile Thr Cys
20 25 30
Val Val Leu Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Gln Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile Glu
65 70 75 80
His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val Asn His
85 90 95
Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys Glu
115 120 125
Leu Ser Ser Ser Asp Thr Val Thr Leu Thr Cys Leu Ile Lys Asp Phe
130 135 140
Phe Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Pro Glu
145 150 155 160
Pro Glu Ser Lys Tyr His Thr Thr Ala Pro Gln Leu Asp Glu Asp Gly
165 170 175
Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln
180 185 190
Gln Gly Asp Thr Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn
195 200 205
His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 46
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q-
K(93)R
<400> 46
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 47
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-C Fc protein A-C1q-
CD16 + K(93)R
<400> 47
Pro Gly Cys Gly Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Ile Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asn Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu
50 55 60
Glu Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Arg Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly
100 105 110
Gln Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu
115 120 125
Met Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe
130 135 140
Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 48
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q +
CD16 - M(5)P
<400> 48
Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 49
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q +
CD16 - P(39)R
<400> 49
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Arg Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 50
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q +
CD16 - D(38)G
<400> 50
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Gly Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 51
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q +
CD16 - D(38)G P(39)R
<400> 51
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 52
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q +
CD16 - K(97)I
<400> 52
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Ile Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 53
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q +
CD16 - A(98)G
<400> 53
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Lys Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 54
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q +
CD16 - D(38)G K(97)I A(98)G
<400> 54
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Gly Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 55
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q +
CD16 - M(5)P P(39)R
<400> 55
Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Arg Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 56
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-C Fc protein A-C1q +
CD16 - L(5)P
<400> 56
Pro Gly Cys Gly Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Ile Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asn Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu
50 55 60
Glu Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly
100 105 110
Gln Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu
115 120 125
Met Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe
130 135 140
Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 57
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-C Fc protein A-C1q +
CD16 - P(39)R
<400> 57
Pro Gly Cys Gly Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Ile Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Arg Glu Asn Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu
50 55 60
Glu Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly
100 105 110
Gln Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu
115 120 125
Met Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe
130 135 140
Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 58
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-C Fc protein A-C1q +
CD16 - D(38)G
<400> 58
Pro Gly Cys Gly Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Ile Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys
20 25 30
Val Val Val Asp Leu Gly Pro Glu Asn Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu
50 55 60
Glu Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly
100 105 110
Gln Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu
115 120 125
Met Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe
130 135 140
Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 59
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-C Fc protein A-C1q +
CD16 - K(97)I
<400> 59
Pro Gly Cys Gly Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Ile Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asn Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu
50 55 60
Glu Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Lys Val Asn Asn
85 90 95
Ile Ala Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly
100 105 110
Gln Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu
115 120 125
Met Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe
130 135 140
Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 60
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-C Fc protein A-C1q +
CD16 - A(98)G
<400> 60
Pro Gly Cys Gly Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Ile Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asn Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu
50 55 60
Glu Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Lys Val Asn Asn
85 90 95
Lys Gly Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly
100 105 110
Gln Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu
115 120 125
Met Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe
130 135 140
Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 61
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-C Fc protein A-C1q +
CD16 - L(5)P P(39)R
<400> 61
Pro Gly Cys Gly Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Ile Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Arg Glu Asn Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu
50 55 60
Glu Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly
100 105 110
Gln Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu
115 120 125
Met Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe
130 135 140
Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 62
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-C Fc protein A-C1q +
CD16 - D(38)G K(97)I A(98)G
<400> 62
Pro Gly Cys Gly Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Ile Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys
20 25 30
Val Val Val Asp Leu Gly Pro Glu Asn Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu
50 55 60
Glu Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Lys Val Asn Asn
85 90 95
Ile Gly Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly
100 105 110
Gln Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu
115 120 125
Met Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe
130 135 140
Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 63
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variants of canine IgG-C Fc C1q-K (93) R
Protein A + I (21) T V (23) L T (24) I
<400> 63
Pro Gly Cys Gly Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Thr Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asn Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu
50 55 60
Glu Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Arg Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly
100 105 110
Gln Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu
115 120 125
Met Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe
130 135 140
Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 64
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q-
CD16 - K(93)R K(97)I A(98)G
<400> 64
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn
85 90 95
Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 65
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q-
CD16 - D(38)G K(93)R K(97)I A(98)G
<400> 65
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Gly Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn
85 90 95
Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 66
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q-
CD16 - M(5)P P(39)R K(93)R
<400> 66
Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Arg Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 67
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q-
CD16 - M(5)P D(38)G P(39)R K(93)R
<400> 67
Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 68
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-C Fc protein A-C1q-
CD16 - D(38)G K(93)R K(97)I A(98)G
<400> 68
Pro Gly Cys Gly Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Ile Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys
20 25 30
Val Val Val Asp Leu Gly Pro Glu Asn Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu
50 55 60
Glu Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Arg Val Asn Asn
85 90 95
Ile Gly Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly
100 105 110
Gln Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu
115 120 125
Met Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe
130 135 140
Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 69
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-C Fc protein A-C1q-
CD16 - M(5)P P(39)R K(93)R
<400> 69
Pro Gly Cys Gly Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Ile Leu Val Thr Ala Arg Thr Pro Thr Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Arg Glu Asn Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Ser Lys Gln Val Gln Thr Ala Asn Thr Gln Pro Arg Glu
50 55 60
Glu Gln Ser Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Ser Gly Lys Gln Phe Lys Cys Arg Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Glu Ile Ile Ser Lys Thr Pro Gly
100 105 110
Gln Ala His Gln Pro Asn Val Tyr Val Leu Pro Pro Ser Arg Asp Glu
115 120 125
Met Ser Lys Asn Thr Val Thr Leu Thr Cys Leu Val Lys Asp Phe Phe
130 135 140
Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Ile Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 70
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type equine IgG1Fc protein A + C1q +
<400> 70
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Asn Pro Lys Asp Thr Leu
1 5 10 15
Met Ile Thr Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
20 25 30
Gln Glu Asn Pro Asp Val Lys Phe Asn Trp Tyr Met Asp Gly Val Glu
35 40 45
Val Arg Thr Ala Thr Thr Arg Pro Lys Glu Glu Gln Phe Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Val Leu Arg Ile Gln His Gln Asp Trp Leu Ser
65 70 75 80
Gly Lys Glu Phe Lys Cys Lys Val Asn Asn Gln Ala Leu Pro Gln Pro
85 90 95
Ile Glu Arg Thr Ile Thr Lys Thr Lys Gly Arg Ser Gln Glu Pro Gln
100 105 110
Val Tyr Val Leu Ala Pro His Pro Asp Glu Ser Lys Lys Ser Lys Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Glu Ile Asn Ile
130 135 140
Glu Trp Gln Ser Asn Gly Gln Pro Glu Leu Glu Thr Lys Tyr Ser Thr
145 150 155 160
Thr Gln Ala Gln Gln Asp Ser Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Ser Val Asp Arg Asn Arg Trp Gln Gln Gly Thr Thr Phe Thr Cys
180 185 190
Gly Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Asn Val
195 200 205
Ser Lys Asn Pro Gly Lys
210
<210> 71
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type equine IgG2Fc protein A-C1q-
<400> 71
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Ala Leu
1 5 10 15
Met Ile Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser
20 25 30
Asp Gln Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu
35 40 45
Val His Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser
65 70 75 80
Gly Lys Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro
85 90 95
Ile Ser Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln
100 105 110
Val Tyr Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val
130 135 140
Glu Trp Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr
145 150 155 160
Thr Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Ser Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys
180 185 190
Ala Val Met His Glu Ala Leu His Asn His Phe Thr Lys Thr Asp Ile
195 200 205
Ser Glu Ser Leu Gly Lys
210
<210> 72
<211> 244
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type equine IgG2Fc protein with hinge
A - C1q -
<400> 72
Pro Pro Cys Val Leu Ser Ala Glu Gly Val Ile Pro Ile Pro Ser Val
1 5 10 15
Pro Lys Pro Gln Cys Pro Pro Tyr Thr His Ser Lys Phe Leu Gly Gly
20 25 30
Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Ala Leu Met Ile
35 40 45
Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln
50 55 60
Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His
65 70 75 80
Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg
85 90 95
Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys
100 105 110
Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser
115 120 125
Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr
130 135 140
Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val
145 150 155 160
Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp
165 170 175
Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro
180 185 190
Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser
195 200 205
Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val
210 215 220
Met His Glu Ala Leu His Asn His Phe Thr Lys Thr Asp Ile Ser Glu
225 230 235 240
Ser Leu Gly Lys
<210> 73
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type equine IgG3 Fc protein A + C1q +
<400> 73
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu
1 5 10 15
Met Ile Thr Arg Met Pro Glu Val Thr Cys Leu Val Val Asp Val Ser
20 25 30
His Asp Ser Ser Asp Val Leu Phe Thr Trp Tyr Val Asp Gly Thr Glu
35 40 45
Val Lys Thr Ala Lys Thr Met Pro Asn Glu Glu Gln Asn Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Val Leu Arg Ile Gln His Gln Asp Trp Leu Asn
65 70 75 80
Gly Lys Lys Phe Lys Cys Lys Val Asn Asn Gln Ala Leu Pro Ala Pro
85 90 95
Val Glu Arg Thr Ile Ser Lys Ala Thr Gly Gln Thr Arg Val Pro Gln
100 105 110
Val Tyr Val Leu Ala Pro His Pro Asp Glu Leu Ser Lys Asn Lys Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Thr Val
130 135 140
Glu Trp Gln Ser Asn Glu His Pro Glu Pro Glu Gly Lys Tyr Arg Thr
145 150 155 160
Thr Glu Ala Gln Lys Asp Ser Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Thr Val Glu Lys Asp Arg Trp Gln Gln Gly Thr Thr Phe Thr Cys
180 185 190
Val Val Met His Glu Ala Leu His Asn His Val Met Gln Lys Asn Ile
195 200 205
Ser Lys Asn Pro Gly Lys
210
<210> 74
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type equine IgG4 Fc protein A + C1q +
<400> 74
Val Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu
1 5 10 15
Met Ile Ser Arg Thr Pro Thr Val Thr Cys Val Val Val Asp Val Gly
20 25 30
His Asp Phe Pro Asp Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu
35 40 45
Thr His Thr Ala Thr Thr Glu Pro Lys Gln Glu Gln Phe Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Val Leu Pro Ile Gln His Lys Asp Trp Leu Ser
65 70 75 80
Gly Lys Glu Phe Lys Cys Lys Val Asn Asn Lys Ala Leu Pro Ala Pro
85 90 95
Val Glu Arg Thr Ile Ser Ala Pro Thr Gly Gln Pro Arg Glu Pro Gln
100 105 110
Val Tyr Val Leu Ala Pro His Arg Asp Glu Leu Ser Lys Asn Lys Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Asp Ile
130 135 140
Glu Trp Lys Ser Asn Gly Gln Pro Glu Pro Glu Thr Lys Tyr Ser Thr
145 150 155 160
Thr Pro Ala Gln Leu Asp Ser Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Thr Val Glu Thr Asn Arg Trp Gln Gln Gly Thr Thr Phe Thr Cys
180 185 190
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Glu Lys Ser Val
195 200 205
Ser Lys Ser Pro Gly Lys
210
<210> 75
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type equine IgG5 Fc protein A-C1q-
<400> 75
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu
1 5 10 15
Met Ile Ser Arg Lys Pro Glu Val Thr Cys Val Val Val Asp Leu Gly
20 25 30
His Asp Asp Pro Asp Val Gln Phe Thr Trp Phe Val Asp Gly Val Glu
35 40 45
Thr His Thr Ala Thr Thr Glu Pro Lys Glu Glu Gln Phe Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser
65 70 75 80
Gly Lys Glu Phe Lys Cys Ser Val Thr Ser Lys Ala Leu Pro Ala Pro
85 90 95
Val Glu Arg Thr Ile Ser Lys Ala Lys Gly Gln Leu Arg Val Pro Gln
100 105 110
Val Tyr Val Leu Ala Pro His Pro Asp Glu Leu Ala Lys Asn Thr Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Glu Ile Asp Val
130 135 140
Glu Trp Gln Ser Asn Glu His Pro Glu Pro Glu Gly Lys Tyr Ser Thr
145 150 155 160
Thr Pro Ala Gln Leu Asn Ser Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Ser Val Glu Thr Ser Arg Trp Lys Gln Gly Glu Ser Phe Thr Cys
180 185 190
Gly Val Met His Glu Ala Val Glu Asn His Tyr Thr Gln Lys Asn Val
195 200 205
Ser His Ser Pro Gly Lys
210
<210> 76
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type equine IgG6Fc protein A-C1q-
<400> 76
Gly Arg Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu
1 5 10 15
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
20 25 30
Gln Glu Asn Pro Asp Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
35 40 45
Ala His Thr Ala Thr Thr Lys Ala Lys Glu Lys Gln Asp Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Arg Arg
65 70 75 80
Gly Lys Glu Phe Lys Cys Lys Val Asn Asn Arg Ala Leu Pro Ala Pro
85 90 95
Val Glu Arg Thr Ile Thr Lys Ala Lys Gly Glu Leu Gln Asp Pro Gln
100 105 110
Val Tyr Ile Leu Ala Pro His Pro Asp Glu Val Thr Lys Asn Thr Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Asn Val
130 135 140
Glu Trp Gln Ser Asn Glu Glu Pro Glu Pro Glu Val Lys Tyr Ser Thr
145 150 155 160
Thr Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Thr Val Glu Thr Asp Arg Trp Glu Gln Gly Glu Ser Phe Thr Cys
180 185 190
Val Val Met His Glu Ala Ile Arg His Thr Tyr Arg Gln Lys Ser Ile
195 200 205
Thr Asn Phe Pro Gly Lys
210
<210> 77
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type equine IgG7Fc protein A + C1q +
<400> 77
Val Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu
1 5 10 15
Met Ile Ser Arg Thr Pro Thr Val Thr Cys Val Val Val Asp Val Gly
20 25 30
His Asp Phe Pro Asp Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu
35 40 45
Thr His Thr Ala Thr Thr Glu Pro Lys Gln Glu Gln Asn Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Ile Leu Ala Ile Gln His Lys Asp Trp Leu Ser
65 70 75 80
Gly Lys Glu Phe Lys Cys Lys Val Asn Asn Gln Ala Leu Pro Ala Pro
85 90 95
Val Gln Lys Thr Ile Ser Lys Pro Thr Gly Gln Pro Arg Glu Pro Gln
100 105 110
Val Tyr Val Leu Ala Pro His Pro Asp Glu Leu Ser Lys Asn Lys Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Asp Ile
130 135 140
Glu Trp Lys Ser Asn Gly Gln Pro Glu Pro Glu Thr Lys Tyr Ser Thr
145 150 155 160
Thr Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Thr Val Glu Thr Asn Arg Trp Gln Gln Gly Thr Thr Phe Thr Cys
180 185 190
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Glu Lys Ser Val
195 200 205
Ser Lys Ser Pro Gly Lys
210
<210> 78
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG2Fc C1 q-protein A +
F(203)Y
<400> 78
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Ala Leu
1 5 10 15
Met Ile Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser
20 25 30
Asp Gln Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu
35 40 45
Val His Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser
65 70 75 80
Gly Lys Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro
85 90 95
Ile Ser Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln
100 105 110
Val Tyr Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val
130 135 140
Glu Trp Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr
145 150 155 160
Thr Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Ser Leu Glu Thr Ser Arg Trp Gln Gln Gly Glu Ser Phe Thr Cys
180 185 190
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Lys Thr Asp Ile
195 200 205
Ser Glu Ser Leu Gly Lys
210
<210> 79
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG2Fc C1 q-protein A +
A(15)T F(203)Y
<400> 79
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu
1 5 10 15
Met Ile Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser
20 25 30
Asp Gln Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu
35 40 45
Val His Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser
65 70 75 80
Gly Lys Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro
85 90 95
Ile Ser Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln
100 105 110
Val Tyr Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val
130 135 140
Glu Trp Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr
145 150 155 160
Thr Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Ser Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys
180 185 190
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Lys Thr Asp Ile
195 200 205
Ser Glu Ser Leu Gly Lys
210
<210> 80
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG5 Fc C1 q-protein A +
V(199)L E(200)H
<400> 80
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu
1 5 10 15
Met Ile Ser Arg Lys Pro Glu Val Thr Cys Val Val Val Asp Leu Gly
20 25 30
His Asp Asp Pro Asp Val Gln Phe Thr Trp Phe Val Asp Gly Val Glu
35 40 45
Thr His Thr Ala Thr Thr Glu Pro Lys Glu Glu Gln Phe Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser
65 70 75 80
Gly Lys Glu Phe Lys Cys Ser Val Thr Ser Lys Ala Leu Pro Ala Pro
85 90 95
Val Glu Arg Thr Ile Ser Lys Ala Lys Gly Gln Leu Arg Val Pro Gln
100 105 110
Val Tyr Val Leu Ala Pro His Pro Asp Glu Leu Ala Lys Asn Thr Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Glu Ile Asp Val
130 135 140
Glu Trp Gln Ser Asn Glu His Pro Glu Pro Glu Gly Lys Tyr Ser Thr
145 150 155 160
Thr Pro Ala Gln Leu Asn Ser Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Ser Val Glu Thr Ser Arg Trp Lys Gln Gly Glu Ser Phe Thr Cys
180 185 190
Gly Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Asn Val
195 200 205
Ser His Ser Pro Gly Lys
210
<210> 81
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG6Fc C1 q-protein A +
I(199)L R(200)H H(201)N T(202)H
<400> 81
Gly Arg Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu
1 5 10 15
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
20 25 30
Gln Glu Asn Pro Asp Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
35 40 45
Ala His Thr Ala Thr Thr Lys Ala Lys Glu Lys Gln Asp Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Arg Arg
65 70 75 80
Gly Lys Glu Phe Lys Cys Lys Val Asn Asn Arg Ala Leu Pro Ala Pro
85 90 95
Val Glu Arg Thr Ile Thr Lys Ala Lys Gly Glu Leu Gln Asp Pro Gln
100 105 110
Val Tyr Ile Leu Ala Pro His Pro Asp Glu Val Thr Lys Asn Thr Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Asn Val
130 135 140
Glu Trp Gln Ser Asn Glu Glu Pro Glu Pro Glu Val Lys Tyr Ser Thr
145 150 155 160
Thr Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Thr Val Glu Thr Asp Arg Trp Glu Gln Gly Glu Ser Phe Thr Cys
180 185 190
Val Val Met His Glu Ala Leu His Asn His Tyr Arg Gln Lys Ser Ile
195 200 205
Thr Asn Phe Pro Gly Lys
210
<210> 82
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG1Fc protein A + C1q-
K(87)S
<400> 82
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Asn Pro Lys Asp Thr Leu
1 5 10 15
Met Ile Thr Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
20 25 30
Gln Glu Asn Pro Asp Val Lys Phe Asn Trp Tyr Met Asp Gly Val Glu
35 40 45
Val Arg Thr Ala Thr Thr Arg Pro Lys Glu Glu Gln Phe Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Val Leu Arg Ile Gln His Gln Asp Trp Leu Ser
65 70 75 80
Gly Lys Glu Phe Lys Cys Ser Val Asn Asn Gln Ala Leu Pro Gln Pro
85 90 95
Ile Glu Arg Thr Ile Thr Lys Thr Lys Gly Arg Ser Gln Glu Pro Gln
100 105 110
Val Tyr Val Leu Ala Pro His Pro Asp Glu Ser Lys Lys Ser Lys Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Glu Ile Asn Ile
130 135 140
Glu Trp Gln Ser Asn Gly Gln Pro Glu Leu Glu Thr Lys Tyr Ser Thr
145 150 155 160
Thr Gln Ala Gln Gln Asp Ser Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Ser Val Asp Arg Asn Arg Trp Gln Gln Gly Thr Thr Phe Thr Cys
180 185 190
Gly Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Asn Val
195 200 205
Ser Lys Asn Pro Gly Lys
210
<210> 83
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG3 Fc protein A + C1q-
K(87)S
<400> 83
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu
1 5 10 15
Met Ile Thr Arg Met Pro Glu Val Thr Cys Leu Val Val Asp Val Ser
20 25 30
His Asp Ser Ser Asp Val Leu Phe Thr Trp Tyr Val Asp Gly Thr Glu
35 40 45
Val Lys Thr Ala Lys Thr Met Pro Asn Glu Glu Gln Asn Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Val Leu Arg Ile Gln His Gln Asp Trp Leu Asn
65 70 75 80
Gly Lys Lys Phe Lys Cys Ser Val Asn Asn Gln Ala Leu Pro Ala Pro
85 90 95
Val Glu Arg Thr Ile Ser Lys Ala Thr Gly Gln Thr Arg Val Pro Gln
100 105 110
Val Tyr Val Leu Ala Pro His Pro Asp Glu Leu Ser Lys Asn Lys Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Thr Val
130 135 140
Glu Trp Gln Ser Asn Glu His Pro Glu Pro Glu Gly Lys Tyr Arg Thr
145 150 155 160
Thr Glu Ala Gln Lys Asp Ser Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Thr Val Glu Lys Asp Arg Trp Gln Gln Gly Thr Thr Phe Thr Cys
180 185 190
Val Val Met His Glu Ala Leu His Asn His Val Met Gln Lys Asn Ile
195 200 205
Ser Lys Asn Pro Gly Lys
210
<210> 84
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG4 Fc protein A + C1q-
K(87)S
<400> 84
Val Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu
1 5 10 15
Met Ile Ser Arg Thr Pro Thr Val Thr Cys Val Val Val Asp Val Gly
20 25 30
His Asp Phe Pro Asp Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu
35 40 45
Thr His Thr Ala Thr Thr Glu Pro Lys Gln Glu Gln Phe Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Val Leu Pro Ile Gln His Lys Asp Trp Leu Ser
65 70 75 80
Gly Lys Glu Phe Lys Cys Ser Val Asn Asn Lys Ala Leu Pro Ala Pro
85 90 95
Val Glu Arg Thr Ile Ser Ala Pro Thr Gly Gln Pro Arg Glu Pro Gln
100 105 110
Val Tyr Val Leu Ala Pro His Arg Asp Glu Leu Ser Lys Asn Lys Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Asp Ile
130 135 140
Glu Trp Lys Ser Asn Gly Gln Pro Glu Pro Glu Thr Lys Tyr Ser Thr
145 150 155 160
Thr Pro Ala Gln Leu Asp Ser Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Thr Val Glu Thr Asn Arg Trp Gln Gln Gly Thr Thr Phe Thr Cys
180 185 190
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Glu Lys Ser Val
195 200 205
Ser Lys Ser Pro Gly Lys
210
<210> 85
<211> 214
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG7Fc protein A + C1q-
K(87)S
<400> 85
Val Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu
1 5 10 15
Met Ile Ser Arg Thr Pro Thr Val Thr Cys Val Val Val Asp Val Gly
20 25 30
His Asp Phe Pro Asp Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu
35 40 45
Thr His Thr Ala Thr Thr Glu Pro Lys Gln Glu Gln Asn Asn Ser Thr
50 55 60
Tyr Arg Val Val Ser Ile Leu Ala Ile Gln His Lys Asp Trp Leu Ser
65 70 75 80
Gly Lys Glu Phe Lys Cys Ser Val Asn Asn Gln Ala Leu Pro Ala Pro
85 90 95
Val Gln Lys Thr Ile Ser Lys Pro Thr Gly Gln Pro Arg Glu Pro Gln
100 105 110
Val Tyr Val Leu Ala Pro His Pro Asp Glu Leu Ser Lys Asn Lys Val
115 120 125
Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Asp Ile
130 135 140
Glu Trp Lys Ser Asn Gly Gln Pro Glu Pro Glu Thr Lys Tyr Ser Thr
145 150 155 160
Thr Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
165 170 175
Leu Thr Val Glu Thr Asn Arg Trp Gln Gln Gly Thr Thr Phe Thr Cys
180 185 190
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Glu Lys Ser Val
195 200 205
Ser Lys Ser Pro Gly Lys
210
<210> 86
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild type cat IgG1a Fc protein A + C1q +
<400> 86
Arg Lys Thr Asp His Pro Pro Gly Pro Lys Thr Gly Glu Gly Pro Lys
1 5 10 15
Cys Pro Pro Pro Glu Met Leu Gly Gly Pro Ser Ile Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asp Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Gln Val Tyr Thr Ala Lys Thr Ser Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Ala Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Ser Phe
145 150 155 160
His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Arg Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Val Tyr Ser Lys Leu Ser Val Asp Arg Ser His Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 87
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild type cat IgG1a Fc protein A + C1q +
<400> 87
Arg Lys Thr Asp His Pro Pro Gly Pro Lys Pro Cys Asp Cys Pro Lys
1 5 10 15
Cys Pro Pro Pro Glu Met Leu Gly Gly Pro Ser Ile Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asp Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Gln Val Tyr Thr Ala Lys Thr Ser Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Ala Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Ser Phe
145 150 155 160
His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Arg Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Val Tyr Ser Lys Leu Ser Val Asp Arg Ser His Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 88
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild type cat IgG1b Fc protein A + C1q +
<400> 88
Arg Lys Thr Asp His Pro Pro Gly Pro Lys Thr Gly Glu Gly Pro Lys
1 5 10 15
Cys Pro Pro Pro Glu Met Leu Gly Gly Pro Ser Ile Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asp Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Gln Val Tyr Thr Ala Lys Thr Ser Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Asp Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Ala Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Glu Gly Phe
145 150 155 160
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Arg Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser Arg Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 89
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild type cat IgG1b Fc protein A + C1q +
<400> 89
Arg Lys Thr Asp His Pro Pro Gly Pro Lys Pro Cys Asp Cys Pro Lys
1 5 10 15
Cys Pro Pro Pro Glu Met Leu Gly Gly Pro Ser Ile Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asp Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Gln Val Tyr Thr Ala Lys Thr Ser Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Asp Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Ala Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Glu Gly Phe
145 150 155 160
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Arg Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser Arg Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 90
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary wild-type cat class IgG2Fc protein A + C1q-
<400> 90
Pro Lys Thr Ala Ser Thr Ile Glu Ser Lys Thr Gly Glu Gly Pro Lys
1 5 10 15
Cys Pro Val Pro Glu Ile Pro Gly Ala Pro Ser Val Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asn Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Glu Met His Thr Ala Lys Thr Arg Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Ala Met Glu Arg Thr Ile Ser Lys Ala Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Thr Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Gly Phe
145 150 155 160
His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Gln Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser His Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 91
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant cat class IgG1a Fc protein A + C1q-
P(198)A
<400> 91
Arg Lys Thr Asp His Pro Pro Gly Pro Lys Pro Cys Asp Cys Pro Lys
1 5 10 15
Cys Pro Pro Pro Glu Met Leu Gly Gly Pro Ser Ile Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asp Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Gln Val Tyr Thr Ala Lys Thr Ser Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Ala Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Ser Phe
145 150 155 160
His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Arg Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Val Tyr Ser Lys Leu Ser Val Asp Arg Ser His Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 92
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant cat class IgG1a Fc protein A + C1q-
P(198)A
<400> 92
Arg Lys Thr Asp His Pro Pro Gly Pro Lys Thr Gly Glu Gly Pro Lys
1 5 10 15
Cys Pro Pro Pro Glu Met Leu Gly Gly Pro Ser Ile Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asp Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Gln Val Tyr Thr Ala Lys Thr Ser Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Ala Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Ser Phe
145 150 155 160
His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Arg Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Val Tyr Ser Lys Leu Ser Val Asp Arg Ser His Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 93
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant cat class IgG1b Fc protein A + C1q-
P(198)A
<400> 93
Arg Lys Thr Asp His Pro Pro Gly Pro Lys Pro Cys Asp Cys Pro Lys
1 5 10 15
Cys Pro Pro Pro Glu Met Leu Gly Gly Pro Ser Ile Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asp Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Gln Val Tyr Thr Ala Lys Thr Ser Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Asp Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Ala Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Glu Gly Phe
145 150 155 160
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Arg Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser Arg Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 94
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant cat class IgG1b Fc protein A + C1q-
P(198)A
<400> 94
Arg Lys Thr Asp His Pro Pro Gly Pro Lys Thr Gly Glu Gly Pro Lys
1 5 10 15
Cys Pro Pro Pro Glu Met Leu Gly Gly Pro Ser Ile Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asp Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Gln Val Tyr Thr Ala Lys Thr Ser Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Asp Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Ala Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Glu Gly Phe
145 150 155 160
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Arg Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser Arg Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 95
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant cat IgG2Fc hinge Cys G (14) C
<400> 95
Pro Lys Thr Ala Ser Thr Ile Glu Ser Lys Thr Gly Glu Cys Pro Lys
1 5 10 15
Cys Pro Val Pro Glu Ile Pro Gly Ala Pro Ser Val Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asn Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Glu Met His Thr Ala Lys Thr Arg Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Ala Met Glu Arg Thr Ile Ser Lys Ala Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Thr Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Gly Phe
145 150 155 160
His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Gln Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser His Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 96
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant cat class IgG1a Fc with modified hinge
K(16)P
<400> 96
Arg Lys Thr Asp His Pro Pro Gly Pro Lys Thr Gly Glu Gly Pro Pro
1 5 10 15
Cys Pro Pro Pro Glu Met Leu Gly Gly Pro Ser Ile Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asp Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Gln Val Tyr Thr Ala Lys Thr Ser Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Ala Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Ser Phe
145 150 155 160
His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Arg Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Val Tyr Ser Lys Leu Ser Val Asp Arg Ser His Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 97
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant cat class IgG1a Fc with modified hinge
K(16)P
<400> 97
Arg Lys Thr Asp His Pro Pro Gly Pro Lys Pro Cys Asp Cys Pro Pro
1 5 10 15
Cys Pro Pro Pro Glu Met Leu Gly Gly Pro Ser Ile Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asp Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Gln Val Tyr Thr Ala Lys Thr Ser Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Ala Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Ser Phe
145 150 155 160
His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Arg Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Val Tyr Ser Lys Leu Ser Val Asp Arg Ser His Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 98
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant cat class IgG1b Fc with modified hinge
K(16)P
<400> 98
Arg Lys Thr Asp His Pro Pro Gly Pro Lys Thr Gly Glu Gly Pro Pro
1 5 10 15
Cys Pro Pro Pro Glu Met Leu Gly Gly Pro Ser Ile Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asp Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Gln Val Tyr Thr Ala Lys Thr Ser Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Asp Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Ala Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Glu Gly Phe
145 150 155 160
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Arg Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser Arg Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 99
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant cat class IgG1b Fc with modified hinge
K(16)P
<400> 99
Arg Lys Thr Asp His Pro Pro Gly Pro Lys Pro Cys Asp Cys Pro Pro
1 5 10 15
Cys Pro Pro Pro Glu Met Leu Gly Gly Pro Ser Ile Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asp Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Gln Val Tyr Thr Ala Lys Thr Ser Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Asp Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Ala Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Glu Gly Phe
145 150 155 160
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Arg Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser Arg Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 100
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant cat class IgG2Fc with modified hinge
K(16)P
<400> 100
Pro Lys Thr Ala Ser Thr Ile Glu Ser Lys Thr Gly Glu Gly Pro Pro
1 5 10 15
Cys Pro Val Pro Glu Ile Pro Gly Ala Pro Ser Val Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asn Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Glu Met His Thr Ala Lys Thr Arg Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Ala Met Glu Arg Thr Ile Ser Lys Ala Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Thr Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Gly Phe
145 150 155 160
His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Gln Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser His Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 101
<211> 244
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG2Fc with modified hinge
Protein A-C1Q-Q (20) P
<400> 101
Pro Pro Cys Val Leu Ser Ala Glu Gly Val Ile Pro Ile Pro Ser Val
1 5 10 15
Pro Lys Pro Pro Cys Pro Pro Tyr Thr His Ser Lys Phe Leu Gly Gly
20 25 30
Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Ala Leu Met Ile
35 40 45
Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln
50 55 60
Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His
65 70 75 80
Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg
85 90 95
Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys
100 105 110
Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser
115 120 125
Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr
130 135 140
Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val
145 150 155 160
Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp
165 170 175
Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro
180 185 190
Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser
195 200 205
Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val
210 215 220
Met His Glu Ala Leu His Asn His Phe Thr Lys Thr Asp Ile Ser Glu
225 230 235 240
Ser Leu Gly Lys
<210> 102
<211> 244
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG2Fc with modified hinge
Protein A-C1q-C (3) S
<400> 102
Pro Pro Ser Val Leu Ser Ala Glu Gly Val Ile Pro Ile Pro Ser Val
1 5 10 15
Pro Lys Pro Gln Cys Pro Pro Tyr Thr His Ser Lys Phe Leu Gly Gly
20 25 30
Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Ala Leu Met Ile
35 40 45
Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln
50 55 60
Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His
65 70 75 80
Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg
85 90 95
Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys
100 105 110
Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser
115 120 125
Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr
130 135 140
Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val
145 150 155 160
Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp
165 170 175
Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro
180 185 190
Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser
195 200 205
Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val
210 215 220
Met His Glu Ala Leu His Asn His Phe Thr Lys Thr Asp Ile Ser Glu
225 230 235 240
Ser Leu Gly Lys
<210> 103
<211> 244
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG2Fc with modified hinge
Protein A-C1q-C (3) S Q (20) P
<400> 103
Pro Pro Ser Val Leu Ser Ala Glu Gly Val Ile Pro Ile Pro Ser Val
1 5 10 15
Pro Lys Pro Pro Cys Pro Pro Tyr Thr His Ser Lys Phe Leu Gly Gly
20 25 30
Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Ala Leu Met Ile
35 40 45
Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln
50 55 60
Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His
65 70 75 80
Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg
85 90 95
Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys
100 105 110
Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser
115 120 125
Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr
130 135 140
Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val
145 150 155 160
Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp
165 170 175
Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro
180 185 190
Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser
195 200 205
Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val
210 215 220
Met His Glu Ala Leu His Asn His Phe Thr Lys Thr Asp Ile Ser Glu
225 230 235 240
Ser Leu Gly Lys
<210> 104
<211> 244
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG2Fc protein A with hinges
+ C1q - A(45)T F(233)Y
<400> 104
Pro Pro Cys Val Leu Ser Ala Glu Gly Val Ile Pro Ile Pro Ser Val
1 5 10 15
Pro Lys Pro Gln Cys Pro Pro Tyr Thr His Ser Lys Phe Leu Gly Gly
20 25 30
Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu Met Ile
35 40 45
Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln
50 55 60
Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His
65 70 75 80
Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg
85 90 95
Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys
100 105 110
Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser
115 120 125
Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr
130 135 140
Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val
145 150 155 160
Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp
165 170 175
Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro
180 185 190
Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser
195 200 205
Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val
210 215 220
Met His Glu Ala Leu His Asn His Tyr Thr Lys Thr Asp Ile Ser Glu
225 230 235 240
Ser Leu Gly Lys
<210> 105
<211> 244
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG2Fc with modified hinge
Protein A + C1Q-Q (20) P A (45) T F (233) Y
<400> 105
Pro Pro Cys Val Leu Ser Ala Glu Gly Val Ile Pro Ile Pro Ser Val
1 5 10 15
Pro Lys Pro Pro Cys Pro Pro Tyr Thr His Ser Lys Phe Leu Gly Gly
20 25 30
Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu Met Ile
35 40 45
Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln
50 55 60
Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His
65 70 75 80
Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg
85 90 95
Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys
100 105 110
Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser
115 120 125
Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr
130 135 140
Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val
145 150 155 160
Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp
165 170 175
Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro
180 185 190
Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser
195 200 205
Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val
210 215 220
Met His Glu Ala Leu His Asn His Tyr Thr Lys Thr Asp Ile Ser Glu
225 230 235 240
Ser Leu Gly Lys
<210> 106
<211> 244
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG2Fc with modified hinge
Protein A + C1q-C (3) S Q (20) P A (45) T F (233) Y
<400> 106
Pro Pro Ser Val Leu Ser Ala Glu Gly Val Ile Pro Ile Pro Ser Val
1 5 10 15
Pro Lys Pro Pro Cys Pro Pro Tyr Thr His Ser Lys Phe Leu Gly Gly
20 25 30
Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu Met Ile
35 40 45
Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln
50 55 60
Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His
65 70 75 80
Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg
85 90 95
Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys
100 105 110
Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser
115 120 125
Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr
130 135 140
Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val
145 150 155 160
Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp
165 170 175
Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro
180 185 190
Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser
195 200 205
Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val
210 215 220
Met His Glu Ala Leu His Asn His Tyr Thr Lys Thr Asp Ile Ser Glu
225 230 235 240
Ser Leu Gly Lys
<210> 107
<211> 237
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Cat IgG2Fc with Cat IgG1 hinge
<400> 107
Arg Lys Thr Asp His Pro Pro Gly Pro Lys Pro Cys Asp Cys Pro Lys
1 5 10 15
Cys Pro Pro Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr
35 40 45
Cys Leu Val Val Asp Leu Gly Pro Asp Asp Ser Asn Val Gln Ile Thr
50 55 60
Trp Phe Val Asp Asn Thr Glu Met His Thr Ala Lys Thr Arg Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Leu His Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn
100 105 110
Ser Lys Ser Leu Pro Ser Ala Met Glu Arg Thr Ile Ser Lys Ala Lys
115 120 125
Gly Gln Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Thr Gln Glu
130 135 140
Glu Leu Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Gly Phe
145 150 155 160
His Pro Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu
165 170 175
Pro Glu Asn Asn Tyr Gln Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly
180 185 190
Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser His Trp Gln
195 200 205
Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser
210 215 220
His His Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
225 230 235
<210> 108
<211> 228
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary variant equine Fc IgG2 (with equine IgG 1)
Hinge) protein A-C1q-
<400> 108
Asp Met Ser Lys Cys Pro Lys Cys Pro Ala Pro Glu Leu Leu Gly Gly
1 5 10 15
Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Ala Leu Met Ile
20 25 30
Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln
35 40 45
Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His
50 55 60
Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg
65 70 75 80
Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys
85 90 95
Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser
100 105 110
Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr
115 120 125
Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val
130 135 140
Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp
145 150 155 160
Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro
165 170 175
Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser
180 185 190
Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val
195 200 205
Met His Glu Ala Leu His Asn His Phe Thr Lys Thr Asp Ile Ser Glu
210 215 220
Ser Leu Gly Lys
225
<210> 109
<211> 228
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant equine IgG2Fc (with equine IgG 1)
Hinge) C1 q-protein A + A (29) T F (217) Y
<400> 109
Asp Met Ser Lys Cys Pro Lys Cys Pro Ala Pro Glu Leu Leu Gly Gly
1 5 10 15
Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu Met Ile
20 25 30
Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln
35 40 45
Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His
50 55 60
Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg
65 70 75 80
Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys
85 90 95
Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser
100 105 110
Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr
115 120 125
Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val
130 135 140
Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp
145 150 155 160
Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro
165 170 175
Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser
180 185 190
Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val
195 200 205
Met His Glu Ala Leu His Asn His Tyr Thr Lys Thr Asp Ile Ser Glu
210 215 220
Ser Leu Gly Lys
225
<210> 110
<211> 384
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-
<400> 110
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro
115 120 125
Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr
130 135 140
Ser Gly Asp Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
145 150 155 160
Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser Val Phe
165 170 175
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
180 185 190
Glu Val Thr Cys Val Val Val Asp Leu Asp Arg Glu Asp Pro Glu Val
195 200 205
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
210 215 220
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
225 230 235 240
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
245 250 255
Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
260 265 270
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
275 280 285
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
290 295 300
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
305 310 315 320
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
325 330 335
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
340 345 350
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
355 360 365
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
370 375 380
<210> 111
<211> 360
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-
<400> 111
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu
130 135 140
Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
145 150 155 160
Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
165 170 175
Leu Asp Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly
180 185 190
Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn
195 200 205
Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp
210 215 220
Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Lys Ala Leu Pro
225 230 235 240
Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln
245 250 255
Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn
260 265 270
Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile
275 280 285
Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr
290 295 300
Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr
305 310 315 320
Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe
325 330 335
Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu
340 345 350
Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 112
<211> 358
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-
<400> 112
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Phe Pro Ala Ser Val Gln Leu His Glu Ala
20 25 30
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe
100 105 110
Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
115 120 125
Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu
130 135 140
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu
145 150 155 160
Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp
165 170 175
Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln
180 185 190
Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr
195 200 205
Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys
210 215 220
Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Lys Ala Leu Pro Ser Pro
225 230 235 240
Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser
245 250 255
Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val
260 265 270
Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val
275 280 285
Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr
290 295 300
Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
305 310 315 320
Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys
325 330 335
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu
340 345 350
Ser His Ser Pro Gly Lys
355
<210> 113
<211> 388
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine with signal sequence
IgG-A Fc C1 q-protein A +
<400> 113
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro
115 120 125
Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr
130 135 140
Ser Gly Asp Ser Gly Gly Gly Ser Gly Gly Gly Ser Phe Asn Glu Cys
145 150 155 160
Arg Cys Thr Asp Thr Pro Cys Pro Val Pro Glu Pro Leu Gly Gly Pro
165 170 175
Ser Val Leu Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala
180 185 190
Arg Thr Pro Glu Val Thr Cys Val Val Leu Asp Leu Gly Arg Glu Asp
195 200 205
Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Glu Val His Thr
210 215 220
Ala Lys Thr Gln Ser Arg Glu Gln Gln Phe Asn Gly Thr Tyr Arg Val
225 230 235 240
Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Thr Gly Lys Glu
245 250 255
Phe Lys Cys Arg Val Asn His Ile Asp Leu Pro Ser Pro Ile Glu Arg
260 265 270
Thr Ile Ser Lys Ala Arg Gly Arg Ala His Lys Pro Ser Val Tyr Val
275 280 285
Leu Pro Pro Ser Pro Lys Glu Leu Ser Ser Ser Asp Thr Val Ser Ile
290 295 300
Thr Cys Leu Ile Lys Asp Phe Tyr Pro Pro Asp Ile Asp Val Glu Trp
305 310 315 320
Gln Ser Asn Gly Gln Gln Glu Pro Glu Arg Lys His Arg Met Thr Pro
325 330 335
Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser
340 345 350
Val Asp Lys Ser Arg Trp Gln Gln Gly Asp Pro Phe Thr Cys Ala Val
355 360 365
Met His Glu Ala Leu His Asn His Tyr Thr Asp Leu Ser Leu Ser His
370 375 380
Ser Pro Gly Lys
385
<210> 114
<211> 364
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine with signal sequence
IgG-A Fc C1 q-protein A +
<400> 114
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Phe Asn Glu Cys Arg Cys Thr Asp Thr Pro Cys Pro
130 135 140
Val Pro Glu Pro Leu Gly Gly Pro Ser Val Leu Ile Phe Pro Pro Lys
145 150 155 160
Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val
165 170 175
Val Leu Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe
180 185 190
Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Ser Arg Glu Gln
195 200 205
Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His
210 215 220
Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val Asn His Ile
225 230 235 240
Asp Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Arg
245 250 255
Ala His Lys Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys Glu Leu
260 265 270
Ser Ser Ser Asp Thr Val Ser Ile Thr Cys Leu Ile Lys Asp Phe Tyr
275 280 285
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
290 295 300
Glu Arg Lys His Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
305 310 315 320
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Gln
325 330 335
Gly Asp Pro Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn His
340 345 350
Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 115
<211> 362
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine with signal sequence
IgG-A Fc C1 q-protein A +
<400> 115
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Phe Pro Ala Ser Val Gln Leu His Glu Ala
20 25 30
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe
100 105 110
Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
115 120 125
Gly Ser Phe Asn Glu Cys Arg Cys Thr Asp Thr Pro Cys Pro Val Pro
130 135 140
Glu Pro Leu Gly Gly Pro Ser Val Leu Ile Phe Pro Pro Lys Pro Lys
145 150 155 160
Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Leu
165 170 175
Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp
180 185 190
Gly Lys Glu Val His Thr Ala Lys Thr Gln Ser Arg Glu Gln Gln Phe
195 200 205
Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp
210 215 220
Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val Asn His Ile Asp Leu
225 230 235 240
Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Arg Ala His
245 250 255
Lys Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys Glu Leu Ser Ser
260 265 270
Ser Asp Thr Val Ser Ile Thr Cys Leu Ile Lys Asp Phe Tyr Pro Pro
275 280 285
Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Arg
290 295 300
Lys His Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe
305 310 315 320
Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Gln Gly Asp
325 330 335
Pro Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr
340 345 350
Asp Leu Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 116
<211> 389
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine with signal sequence
IgG-D Fc C1 q-protein A +
<400> 116
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro
115 120 125
Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr
130 135 140
Ser Gly Asp Ser Gly Gly Gly Ser Gly Gly Gly Ser Pro Lys Glu Ser
145 150 155 160
Thr Cys Lys Cys Ile Ser Pro Cys Pro Val Pro Glu Ser Leu Gly Gly
165 170 175
Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile
180 185 190
Ala Arg Thr Pro Glu Ile Thr Cys Val Val Leu Asp Leu Gly Arg Glu
195 200 205
Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Glu Val His
210 215 220
Thr Ala Lys Thr Gln Pro Arg Glu Gln Gln Phe Asn Ser Thr Tyr Arg
225 230 235 240
Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Thr Gly Lys
245 250 255
Glu Phe Lys Cys Arg Val Asn His Ile Gly Leu Pro Ser Pro Ile Glu
260 265 270
Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr
275 280 285
Val Leu Pro Pro Ser Pro Lys Glu Leu Ser Ser Ser Asp Thr Val Thr
290 295 300
Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Glu Ile Asp Val Glu
305 310 315 320
Trp Gln Ser Asn Gly Gln Pro Glu Pro Glu Ser Lys Tyr His Thr Thr
325 330 335
Ala Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu
340 345 350
Ser Val Asp Lys Ser Arg Trp Gln Gln Gly Asp Thr Phe Thr Cys Ala
355 360 365
Val Met His Glu Ala Leu His Asn His Tyr Thr Asp Leu Ser Leu Ser
370 375 380
His Ser Pro Gly Lys
385
<210> 117
<211> 365
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine with signal sequence
IgG-D Fc C1 q-protein A +
<400> 117
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Pro Lys Glu Ser Thr Cys Lys Cys Ile Ser Pro Cys
130 135 140
Pro Val Pro Glu Ser Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
145 150 155 160
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Ile Thr Cys
165 170 175
Val Val Leu Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp
180 185 190
Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Pro Arg Glu
195 200 205
Gln Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
210 215 220
His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val Asn His
225 230 235 240
Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
245 250 255
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys Glu
260 265 270
Leu Ser Ser Ser Asp Thr Val Thr Leu Thr Cys Leu Ile Lys Asp Phe
275 280 285
Phe Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Pro Glu
290 295 300
Pro Glu Ser Lys Tyr His Thr Thr Ala Pro Gln Leu Asp Glu Asp Gly
305 310 315 320
Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln
325 330 335
Gln Gly Asp Thr Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn
340 345 350
His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
355 360 365
<210> 118
<211> 363
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine with signal sequence
IgG-D Fc C1 q-protein A +
<400> 118
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Phe Pro Ala Ser Val Gln Leu His Glu Ala
20 25 30
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe
100 105 110
Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
115 120 125
Gly Ser Pro Lys Glu Ser Thr Cys Lys Cys Ile Ser Pro Cys Pro Val
130 135 140
Pro Glu Ser Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro
145 150 155 160
Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Ile Thr Cys Val Val
165 170 175
Leu Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val
180 185 190
Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Pro Arg Glu Gln Gln
195 200 205
Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln
210 215 220
Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val Asn His Ile Gly
225 230 235 240
Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala
245 250 255
His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys Glu Leu Ser
260 265 270
Ser Ser Asp Thr Val Thr Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro
275 280 285
Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Pro Glu Pro Glu
290 295 300
Ser Lys Tyr His Thr Thr Ala Pro Gln Leu Asp Glu Asp Gly Ser Tyr
305 310 315 320
Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Gln Gly
325 330 335
Asp Thr Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn His Tyr
340 345 350
Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 119
<211> 373
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD v2 with Signal sequence-with IgG1
Variant horse class IgG 2C 1 q-protein A + of hinge
<400> 119
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Val Ser Phe Pro Ala Ser Val His Leu Gln Thr Ala
20 25 30
Val Glu Gln His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Thr Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Ser Ala Ala Asn Glu Thr Met
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser
100 105 110
Val Met Val Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp
115 120 125
Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Arg
130 135 140
Asp Asp Met Ser Lys Cys Pro Lys Cys Pro Ala Pro Glu Leu Leu Gly
145 150 155 160
Gly Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu Met
165 170 175
Ile Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp
180 185 190
Gln Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val
195 200 205
His Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr
210 215 220
Arg Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly
225 230 235 240
Lys Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile
245 250 255
Ser Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val
260 265 270
Tyr Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser
275 280 285
Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu
290 295 300
Trp Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr
305 310 315 320
Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu
325 330 335
Ser Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala
340 345 350
Val Met His Glu Ala Leu His Asn His Tyr Thr Lys Thr Asp Ile Ser
355 360 365
Glu Ser Leu Gly Lys
370
<210> 120
<211> 349
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD v3 with Signal sequence-with IgG1
Variant horse class IgG 2C 1 q-protein A + of hinge
<400> 120
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Val Ser Phe Pro Ala Ser Val His Leu Gln Thr Ala
20 25 30
Val Glu Gln His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Thr Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Ser Ala Ala Asn Glu Thr Met
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser
100 105 110
Val Met Val Ala Phe Met Asp Asn Pro Asp Met Ser Lys Cys Pro Lys
115 120 125
Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro
130 135 140
Pro Asn Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Val Val Thr
145 150 155 160
Cys Val Val Val Asn Leu Ser Asp Gln Tyr Pro Asp Val Gln Phe Ser
165 170 175
Trp Tyr Val Asp Asn Thr Glu Val His Ser Ala Ile Thr Lys Gln Arg
180 185 190
Glu Ala Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
195 200 205
Gln His Gln Asp Trp Leu Ser Gly Lys Glu Phe Lys Cys Ser Val Thr
210 215 220
Asn Val Gly Val Pro Gln Pro Ile Ser Arg Ala Ile Ser Arg Gly Lys
225 230 235 240
Gly Pro Ser Arg Val Pro Gln Val Tyr Val Leu Pro Pro His Pro Asp
245 250 255
Glu Leu Ala Lys Ser Lys Val Ser Val Thr Cys Leu Val Lys Asp Phe
260 265 270
Tyr Pro Pro Asp Ile Ser Val Glu Trp Gln Ser Asn Arg Trp Pro Glu
275 280 285
Leu Glu Gly Lys Tyr Ser Thr Thr Pro Ala Gln Leu Asp Gly Asp Gly
290 295 300
Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Leu Glu Thr Ser Arg Trp Gln
305 310 315 320
Gln Val Glu Ser Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn
325 330 335
His Tyr Thr Lys Thr Asp Ile Ser Glu Ser Leu Gly Lys
340 345
<210> 121
<211> 347
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD v4 with Signal sequence-with IgG1
Variant horse class IgG 2C 1 q-protein A + of hinge
<400> 121
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Phe Pro Ala Ser Val His Leu Gln Thr Ala Val Glu
20 25 30
Gln His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro
35 40 45
Thr Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe
50 55 60
Ile Phe Thr Glu Phe Leu Glu Ser Ala Ala Asn Glu Thr Met Arg His
65 70 75 80
Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr
85 90 95
Thr Leu Leu Ala Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser Val Met
100 105 110
Val Ala Phe Met Asp Asn Pro Asp Met Ser Lys Cys Pro Lys Cys Pro
115 120 125
Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Asn
130 135 140
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Val Val Thr Cys Val
145 150 155 160
Val Val Asn Leu Ser Asp Gln Tyr Pro Asp Val Gln Phe Ser Trp Tyr
165 170 175
Val Asp Asn Thr Glu Val His Ser Ala Ile Thr Lys Gln Arg Glu Ala
180 185 190
Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gln His
195 200 205
Gln Asp Trp Leu Ser Gly Lys Glu Phe Lys Cys Ser Val Thr Asn Val
210 215 220
Gly Val Pro Gln Pro Ile Ser Arg Ala Ile Ser Arg Gly Lys Gly Pro
225 230 235 240
Ser Arg Val Pro Gln Val Tyr Val Leu Pro Pro His Pro Asp Glu Leu
245 250 255
Ala Lys Ser Lys Val Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro
260 265 270
Pro Asp Ile Ser Val Glu Trp Gln Ser Asn Arg Trp Pro Glu Leu Glu
275 280 285
Gly Lys Tyr Ser Thr Thr Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr
290 295 300
Phe Leu Tyr Ser Lys Leu Ser Leu Glu Thr Ser Arg Trp Gln Gln Val
305 310 315 320
Glu Ser Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn His Tyr
325 330 335
Thr Lys Thr Asp Ile Ser Glu Ser Leu Gly Lys
340 345
<210> 122
<211> 384
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine with signal sequence
IgG-B Fc additional disulfide protein A + C1q-CD16-
<400> 122
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Cys Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Cys Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro
115 120 125
Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr
130 135 140
Ser Gly Asp Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
145 150 155 160
Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser Val Phe
165 170 175
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
180 185 190
Glu Val Thr Cys Val Val Val Asp Leu Asp Arg Glu Asp Pro Glu Val
195 200 205
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
210 215 220
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
225 230 235 240
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
245 250 255
Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
260 265 270
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
275 280 285
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
290 295 300
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
305 310 315 320
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
325 330 335
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
340 345 350
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
355 360 365
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
370 375 380
<210> 123
<211> 360
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine with signal sequence
IgG-B Fc additional disulfide protein A + C1q-CD16-
<400> 123
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Cys Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Cys Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu
130 135 140
Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
145 150 155 160
Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
165 170 175
Leu Asp Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly
180 185 190
Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn
195 200 205
Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp
210 215 220
Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Lys Ala Leu Pro
225 230 235 240
Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln
245 250 255
Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn
260 265 270
Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile
275 280 285
Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr
290 295 300
Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr
305 310 315 320
Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe
325 330 335
Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu
340 345 350
Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 124
<211> 358
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine with signal sequence
IgG-B Fc additional disulfide protein A + C1q-CD16-
<400> 124
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Phe Pro Ala Ser Cys Gln Leu His Glu Ala
20 25 30
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Cys Ala Ala Phe
100 105 110
Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
115 120 125
Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu
130 135 140
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu
145 150 155 160
Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp
165 170 175
Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln
180 185 190
Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr
195 200 205
Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys
210 215 220
Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Lys Ala Leu Pro Ser Pro
225 230 235 240
Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser
245 250 255
Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val
260 265 270
Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val
275 280 285
Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr
290 295 300
Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
305 310 315 320
Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys
325 330 335
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu
340 345 350
Ser His Ser Pro Gly Lys
355
<210> 125
<211> 384
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-
<400> 125
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro
115 120 125
Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr
130 135 140
Ser Gly Asp Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
145 150 155 160
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
165 170 175
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
180 185 190
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
195 200 205
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
210 215 220
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
225 230 235 240
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
245 250 255
Arg Val Asn Asn Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
260 265 270
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
275 280 285
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
290 295 300
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
305 310 315 320
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
325 330 335
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
340 345 350
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
355 360 365
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
370 375 380
<210> 126
<211> 360
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-
<400> 126
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu
130 135 140
Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
145 150 155 160
Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
165 170 175
Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly
180 185 190
Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn
195 200 205
Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp
210 215 220
Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Ile Gly Leu Pro
225 230 235 240
Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln
245 250 255
Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn
260 265 270
Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile
275 280 285
Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr
290 295 300
Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr
305 310 315 320
Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe
325 330 335
Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu
340 345 350
Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 127
<211> 358
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-
<400> 127
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Phe Pro Ala Ser Val Gln Leu His Glu Ala
20 25 30
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe
100 105 110
Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
115 120 125
Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu
130 135 140
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu
145 150 155 160
Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp
165 170 175
Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln
180 185 190
Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr
195 200 205
Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys
210 215 220
Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Ile Gly Leu Pro Ser Pro
225 230 235 240
Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser
245 250 255
Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val
260 265 270
Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val
275 280 285
Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr
290 295 300
Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
305 310 315 320
Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys
325 330 335
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu
340 345 350
Ser His Ser Pro Gly Lys
355
<210> 128
<211> 384
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-
<400> 128
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro
115 120 125
Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr
130 135 140
Ser Gly Asp Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
145 150 155 160
Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser Val Phe
165 170 175
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
180 185 190
Glu Val Thr Cys Val Val Val Asp Leu Gly Arg Glu Asp Pro Glu Val
195 200 205
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
210 215 220
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
225 230 235 240
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
245 250 255
Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
260 265 270
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
275 280 285
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
290 295 300
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
305 310 315 320
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
325 330 335
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
340 345 350
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
355 360 365
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
370 375 380
<210> 129
<211> 360
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-
<400> 129
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu
130 135 140
Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
145 150 155 160
Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
165 170 175
Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly
180 185 190
Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn
195 200 205
Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp
210 215 220
Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Lys Ala Leu Pro
225 230 235 240
Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln
245 250 255
Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn
260 265 270
Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile
275 280 285
Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr
290 295 300
Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr
305 310 315 320
Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe
325 330 335
Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu
340 345 350
Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 130
<211> 358
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-
<400> 130
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Phe Pro Ala Ser Val Gln Leu His Glu Ala
20 25 30
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe
100 105 110
Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
115 120 125
Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu
130 135 140
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu
145 150 155 160
Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Gly
165 170 175
Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln
180 185 190
Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr
195 200 205
Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys
210 215 220
Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Lys Ala Leu Pro Ser Pro
225 230 235 240
Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser
245 250 255
Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val
260 265 270
Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val
275 280 285
Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr
290 295 300
Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
305 310 315 320
Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys
325 330 335
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu
340 345 350
Ser His Ser Pro Gly Lys
355
<210> 131
<211> 470
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3x 2-variant canines with signal sequence
IgG-B Fc protein A + C1q-CD16-
<400> 131
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala
130 135 140
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
145 150 155 160
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
165 170 175
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
180 185 190
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
195 200 205
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe
210 215 220
Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
225 230 235 240
Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu
245 250 255
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu
260 265 270
Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Gly
275 280 285
Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln
290 295 300
Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr
305 310 315 320
Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys
325 330 335
Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Lys Ala Leu Pro Ser Pro
340 345 350
Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser
355 360 365
Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val
370 375 380
Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val
385 390 395 400
Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr
405 410 415
Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
420 425 430
Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys
435 440 445
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu
450 455 460
Ser His Ser Pro Gly Lys
465 470
<210> 132
<211> 367
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant canine IgG-B Fc-canines with Signal sequences
TrkA ECD v3 protein A + C1q-CD16-
<400> 132
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Pro Lys Arg Glu Asn Gly Arg Val Pro Arg
20 25 30
Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly Gly Pro
35 40 45
Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala
50 55 60
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Gly Arg Glu Asp
65 70 75 80
Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr
85 90 95
Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val
100 105 110
Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln
115 120 125
Phe Thr Cys Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg
130 135 140
Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val
145 150 155 160
Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr
165 170 175
Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln
180 185 190
Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro
195 200 205
Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val
210 215 220
Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met
225 230 235 240
His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser
245 250 255
Pro Gly Lys Ser Gly Gly Gly Ser Gly Gly Val Ser Phe Pro Ala Ser
260 265 270
Val Gln Leu His Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe
275 280 285
Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly
290 295 300
Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro
305 310 315 320
Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro
325 330 335
Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser
340 345 350
Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro
355 360 365
<210> 133
<211> 468
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine with signal sequence
IgG-B Fc-Canine TrkA ECD v3 protein A + C1q-CD16-
<400> 133
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu
130 135 140
Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
145 150 155 160
Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
165 170 175
Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly
180 185 190
Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn
195 200 205
Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp
210 215 220
Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Lys Ala Leu Pro
225 230 235 240
Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln
245 250 255
Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn
260 265 270
Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile
275 280 285
Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr
290 295 300
Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr
305 310 315 320
Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe
325 330 335
Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu
340 345 350
Ser Leu Ser His Ser Pro Gly Lys Ser Gly Gly Gly Ser Gly Gly Val
355 360 365
Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His
370 375 380
Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg
385 390 395 400
Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr
405 410 415
Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu
420 425 430
Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu
435 440 445
Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe
450 455 460
Met Asp Asn Pro
465
<210> 134
<211> 541
<212> PRT
<213> domestic dog
<220>
<221> features not yet classified
<223> Canine NGFR
<400> 134
Met Ala Ile Ala Gly Asp Asn Arg Val Ala Val Thr Val Ala Met Gly
1 5 10 15
Arg Ile Val Gly Val Val Ala Met Ala Val Val Ala Val Val Val Leu
20 25 30
Val Val Met Pro Val Ser Trp Leu Arg Gly Gly Arg Ala Arg Phe Arg
35 40 45
Gly Trp Ala Gly Trp Ala Gly Arg Arg Arg Gly Arg Arg Thr Gly Phe
50 55 60
Ser Gln Pro Leu Pro Pro Ser Ala Arg Ala Ser Gly Ala Ser Ala Ser
65 70 75 80
Ser Gly Gly Arg Ala Leu Glu Arg Ser Ala Ala Gln Pro Tyr Pro Ser
85 90 95
Ala Glu Arg Thr Pro Leu Glu Ala Glu Arg Cys His Arg Arg Arg Ala
100 105 110
Val Gly Ala Gly Ala Ala Gly Cys Ala Met Asp Gly Pro Arg Leu Leu
115 120 125
Leu Leu Leu Leu Leu Leu Leu Gly Val Ser Leu Gly Gly Ala Lys Glu
130 135 140
Ala Cys Pro Thr Gly Leu Tyr Thr His Ser Gly Glu Cys Cys Lys Ala
145 150 155 160
Cys Asn Leu Gly Glu Gly Val Ala Gln Pro Cys Gly Ala Asn Gln Thr
165 170 175
Val Cys Glu Pro Cys Leu Asp Ser Val Thr Phe Ser Asp Val Val Ser
180 185 190
Ala Thr Glu Pro Cys Lys Pro Cys Thr Glu Cys Val Gly Leu Gln Ser
195 200 205
Met Ser Ala Pro Cys Val Glu Ala Asp Asp Ala Val Cys Arg Cys Ala
210 215 220
Tyr Gly Tyr Tyr Gln Asp Glu Thr Thr Gly Arg Cys Glu Ala Cys Arg
225 230 235 240
Val Cys Glu Ala Gly Ser Gly Leu Val Phe Ser Cys Gln Asp Arg Gln
245 250 255
Asn Thr Val Cys Glu Glu Cys Pro Asp Gly Thr Tyr Ser Asp Glu Ala
260 265 270
Asn His Val Asp Pro Cys Leu Pro Cys Thr Val Cys Glu Asp Thr Glu
275 280 285
Arg Gln Leu Arg Glu Cys Thr Arg Trp Ala Asp Ala Glu Cys Glu Glu
290 295 300
Ile Pro Gly Arg Trp Ile Thr Arg Ser Thr Pro Ser Glu Asp Ser Asp
305 310 315 320
Ser Thr Ala Pro Ser Thr Glu Glu Pro Glu Leu Pro Pro Asp Gln Glu
325 330 335
Ile Ile Ala Ser Thr Met Ala Asp Val Val Thr Thr Val Met Gly Ser
340 345 350
Ser Gln Pro Val Val Thr Arg Gly Thr Ala Asp Asn Leu Ile Pro Val
355 360 365
Tyr Cys Ser Ile Leu Ala Ala Val Val Val Gly Leu Val Ala Tyr Ile
370 375 380
Ala Phe Lys Arg Trp Asn Ser Cys Lys Gln Asn Lys Gln Gly Ala Asn
385 390 395 400
Ser Arg Pro Val Asn Gln Thr Pro Pro Pro Glu Gly Glu Lys Leu His
405 410 415
Ser Asp Ser Gly Ile Ser Val Asp Ser Gln Ser Leu His Asp Gln Gln
420 425 430
Pro His Thr Gln Thr Ala Ala Gly Gln Ala Leu Lys Gly Asp Gly Gly
435 440 445
Leu Tyr Ser Ser Leu Pro Pro Ala Lys Arg Glu Glu Val Glu Lys Leu
450 455 460
Leu Asn Gly Ser Ala Gly Asp Thr Trp Arg His Leu Ala Gly Glu Leu
465 470 475 480
Gly Tyr Gln Pro Glu His Ile Asp Ser Phe Thr His Glu Ala Cys Pro
485 490 495
Ala Arg Ala Leu Leu Ala Ser Trp Ala Ala Gln Asp Ser Ala Thr Leu
500 505 510
Asp Ala Leu Leu Ala Ala Leu Arg Arg Ile Gln Arg Ala Asp Ile Val
515 520 525
Glu Ser Leu Cys Ser Glu Ser Thr Ala Thr Ser Pro Val
530 535 540
<210> 135
<211> 163
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Canine NGFR ECD
<400> 135
Ala Lys Glu Ala Cys Pro Thr Gly Leu Tyr Thr His Ser Gly Glu Cys
1 5 10 15
Cys Lys Ala Cys Asn Leu Gly Glu Gly Val Ala Gln Pro Cys Gly Ala
20 25 30
Asn Gln Thr Val Cys Glu Pro Cys Leu Asp Ser Val Thr Phe Ser Asp
35 40 45
Val Val Ser Ala Thr Glu Pro Cys Lys Pro Cys Thr Glu Cys Val Gly
50 55 60
Leu Gln Ser Met Ser Ala Pro Cys Val Glu Ala Asp Asp Ala Val Cys
65 70 75 80
Arg Cys Ala Tyr Gly Tyr Tyr Gln Asp Glu Thr Thr Gly Arg Cys Glu
85 90 95
Ala Cys Arg Val Cys Glu Ala Gly Ser Gly Leu Val Phe Ser Cys Gln
100 105 110
Asp Arg Gln Asn Thr Val Cys Glu Glu Cys Pro Asp Gly Thr Tyr Ser
115 120 125
Asp Glu Ala Asn His Val Asp Pro Cys Leu Pro Cys Thr Val Cys Glu
130 135 140
Asp Thr Glu Arg Gln Leu Arg Glu Cys Thr Arg Trp Ala Asp Ala Glu
145 150 155 160
Cys Glu Glu
<210> 136
<211> 430
<212> PRT
<213> domestic cat
<220>
<221> features not yet classified
<223> Cat NGFR
<400> 136
Met Gly Ala Gly Ala Ala Gly Arg Ala Met Asp Gly Pro Arg Pro Leu
1 5 10 15
Leu Leu Leu Leu Pro Leu Leu Leu Gly Val Ser Leu Gly Gly Ala Lys
20 25 30
Glu Ala Cys Pro Thr Gly Leu Phe Thr His Ser Gly Glu Cys Cys Lys
35 40 45
Ala Cys Asn Leu Gly Glu Gly Val Ala Gln Pro Cys Gly Ala Asn Gln
50 55 60
Thr Val Cys Glu Pro Cys Leu Asp Ser Val Thr Phe Ser Asp Val Val
65 70 75 80
Ser Ala Thr Glu Pro Cys Lys Pro Cys Thr Glu Cys Val Gly Leu Gln
85 90 95
Ser Met Ser Ala Pro Cys Val Glu Ala Asp Asp Ala Val Cys Arg Cys
100 105 110
Ala Tyr Gly Tyr Tyr Gln Asp Glu Thr Thr Gly Arg Cys Glu Ala Cys
115 120 125
Arg Val Cys Glu Ala Gly Ser Gly Leu Val Phe Ser Cys Gln Asp Arg
130 135 140
Gln Asn Thr Val Cys Glu Glu Cys Pro Asp Gly Thr Tyr Ser Asp Glu
145 150 155 160
Ala Asn His Val Asp Pro Cys Leu Pro Cys Thr Val Cys Glu Asp Thr
165 170 175
Glu Arg Gln Leu Arg Glu Cys Thr Arg Trp Ala Asp Ala Glu Cys Glu
180 185 190
Glu Ile Pro Gly Arg Trp Ile Thr Arg Ser Thr Pro Ser Glu Gly Ser
195 200 205
Asp Ser Thr Ala Pro Ser Thr Glu Glu Pro Glu Val Pro Pro Glu Gln
210 215 220
Asp Leu Ile Ala Ser Thr Val Ala Asp Val Val Thr Thr Val Met Gly
225 230 235 240
Ser Ser Gln Pro Val Val Thr Arg Gly Thr Ala Asp Asn Leu Ile Pro
245 250 255
Val Tyr Cys Ser Ile Leu Ala Ala Val Val Val Gly Leu Val Ala Tyr
260 265 270
Ile Ala Phe Lys Arg Trp Asn Ser Cys Lys Gln Asp Lys Gln Gly Ala
275 280 285
Asn Ser Arg Pro Val Asn Gln Thr Pro Pro Pro Glu Gly Glu Lys Leu
290 295 300
His Ser Asp Ser Gly Ile Ser Val Asp Ser Gln Ser Leu His Asp Gln
305 310 315 320
Gln Ser His Thr Gln Thr Ala Ala Gly Gln Ala Leu Lys Gly Asp Gly
325 330 335
Gly Leu Tyr Ser Ser Leu Pro Ser Ala Lys Arg Glu Glu Val Glu Lys
340 345 350
Leu Leu Asn Gly Ser Ala Gly Asp Thr Trp Arg His Leu Ala Gly Glu
355 360 365
Leu Gly Tyr Gln Pro Glu His Ile Asp Ser Phe Thr Arg Glu Ala Cys
370 375 380
Pro Ala Arg Ala Leu Leu Ala Ser Trp Ala Ala Gln Asp Ser Ala Thr
385 390 395 400
Leu Asp Ala Leu Leu Ala Ala Leu Arg Arg Ile Gln Arg Ala Asp Ile
405 410 415
Val Glu Ser Leu Cys Ser Glu Ser Thr Ala Thr Ser Pro Val
420 425 430
<210> 137
<211> 163
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Cat NGFR ECD
<400> 137
Ala Lys Glu Ala Cys Pro Thr Gly Leu Phe Thr His Ser Gly Glu Cys
1 5 10 15
Cys Lys Ala Cys Asn Leu Gly Glu Gly Val Ala Gln Pro Cys Gly Ala
20 25 30
Asn Gln Thr Val Cys Glu Pro Cys Leu Asp Ser Val Thr Phe Ser Asp
35 40 45
Val Val Ser Ala Thr Glu Pro Cys Lys Pro Cys Thr Glu Cys Val Gly
50 55 60
Leu Gln Ser Met Ser Ala Pro Cys Val Glu Ala Asp Asp Ala Val Cys
65 70 75 80
Arg Cys Ala Tyr Gly Tyr Tyr Gln Asp Glu Thr Thr Gly Arg Cys Glu
85 90 95
Ala Cys Arg Val Cys Glu Ala Gly Ser Gly Leu Val Phe Ser Cys Gln
100 105 110
Asp Arg Gln Asn Thr Val Cys Glu Glu Cys Pro Asp Gly Thr Tyr Ser
115 120 125
Asp Glu Ala Asn His Val Asp Pro Cys Leu Pro Cys Thr Val Cys Glu
130 135 140
Asp Thr Glu Arg Gln Leu Arg Glu Cys Thr Arg Trp Ala Asp Ala Glu
145 150 155 160
Cys Glu Glu
<210> 138
<211> 429
<212> PRT
<213> horse
<220>
<221> features not yet classified
<223> horse type NGFR
<400> 138
Met Arg Ala Gly Ala Ala Asp Cys Ala Met Asp Gly Pro Arg Leu Leu
1 5 10 15
Leu Leu Leu Leu Leu Leu Gly Val Cys Leu Leu Gly Gly Ala Lys Glu
20 25 30
Val Cys Pro Thr Asp Leu Tyr Thr His Ser Gly Glu Cys Cys Lys Ala
35 40 45
Cys Asn Leu Gly Glu Gly Val Ala Gln Pro Cys Gly Ala Asn Gln Thr
50 55 60
Val Cys Glu Pro Cys Leu Asp Ser Val Thr Phe Ser Asp Val Val Ser
65 70 75 80
Ala Thr Glu Pro Cys Lys Pro Cys Thr Glu Cys Val Gly Leu Gln Ser
85 90 95
Met Ser Ala Pro Cys Val Glu Ala Asp Asp Ala Val Cys Arg Cys Ala
100 105 110
Tyr Gly Tyr Tyr Gln Asp Glu Thr Thr Gly Arg Cys Glu Ala Cys Gln
115 120 125
Val Cys Glu Ala Gly Ser Gly Leu Val Phe Ser Cys Gln Asp Lys Gln
130 135 140
Asn Thr Val Cys Glu Glu Cys Pro Asp Gly Thr Tyr Ser Asp Glu Ala
145 150 155 160
Asn His Val Asp Pro Cys Leu Pro Cys Thr Val Cys Glu Asp Thr Glu
165 170 175
Arg Gln Leu Arg Glu Cys Thr Arg Trp Ala Asp Ala Glu Cys Glu Glu
180 185 190
Ile Pro Ser Arg Trp Ile Thr Arg Ala Thr Pro Pro Glu Gly Ser Asp
195 200 205
Ser Thr Ala Pro Ser Thr Gln Glu Pro Glu Gly Pro Pro Glu Lys Asp
210 215 220
Leu Val Ala Ser Thr Val Ala Asp Val Val Thr Thr Val Met Gly Ser
225 230 235 240
Ser Gln Pro Val Val Thr Arg Gly Thr Thr Asp Asn Leu Ile Pro Val
245 250 255
Tyr Cys Ser Ile Leu Ala Ala Val Val Val Gly Leu Val Ala Tyr Ile
260 265 270
Ala Phe Lys Arg Trp Asn Ser Cys Lys Gln Asn Lys Gln Gly Ala Asn
275 280 285
Ser Arg Pro Val Asn Gln Thr Pro Pro Pro Glu Gly Glu Lys Leu His
290 295 300
Ser Asp Ser Gly Ile Ser Val Asp Ser Gln Ser Leu His Asp Gln Gln
305 310 315 320
Pro His Thr Gln Thr Ala Ala Gly Gln Ala Leu Lys Gly Asp Gly Gly
325 330 335
Leu Tyr Ser Ser Leu Pro Leu Ala Lys Arg Glu Glu Val Glu Lys Leu
340 345 350
Leu Asn Gly Ser Ala Gly Asp Thr Trp Arg His Leu Ala Gly Glu Leu
355 360 365
Gly Tyr Gln Pro Glu His Ile Asp Ser Phe Thr His Glu Ala Cys Pro
370 375 380
Val Arg Ala Leu Leu Ala Ser Trp Ala Ala Gln Asp Ser Ala Thr Phe
385 390 395 400
Asp Ala Leu Leu Thr Ala Leu Arg Arg Ile Gln Arg Ala Asp Ile Val
405 410 415
Glu Ser Leu Cys Ser Glu Ser Thr Ala Thr Ser Pro Val
420 425
<210> 139
<211> 163
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine NGFR ECD
<400> 139
Ala Lys Glu Val Cys Pro Thr Asp Leu Tyr Thr His Ser Gly Glu Cys
1 5 10 15
Cys Lys Ala Cys Asn Leu Gly Glu Gly Val Ala Gln Pro Cys Gly Ala
20 25 30
Asn Gln Thr Val Cys Glu Pro Cys Leu Asp Ser Val Thr Phe Ser Asp
35 40 45
Val Val Ser Ala Thr Glu Pro Cys Lys Pro Cys Thr Glu Cys Val Gly
50 55 60
Leu Gln Ser Met Ser Ala Pro Cys Val Glu Ala Asp Asp Ala Val Cys
65 70 75 80
Arg Cys Ala Tyr Gly Tyr Tyr Gln Asp Glu Thr Thr Gly Arg Cys Glu
85 90 95
Ala Cys Gln Val Cys Glu Ala Gly Ser Gly Leu Val Phe Ser Cys Gln
100 105 110
Asp Lys Gln Asn Thr Val Cys Glu Glu Cys Pro Asp Gly Thr Tyr Ser
115 120 125
Asp Glu Ala Asn His Val Asp Pro Cys Leu Pro Cys Thr Val Cys Glu
130 135 140
Asp Thr Glu Arg Gln Leu Arg Glu Cys Thr Arg Trp Ala Asp Ala Glu
145 150 155 160
Cys Glu Glu
<210> 140
<211> 631
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-canine NGFR ECD-
Variant canine IgG-B Fc-canine TrkA ECD v3
Protein A + C1q-CD16-
<400> 140
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Ala Lys Glu Ala Cys Pro Thr Gly Leu Tyr Thr His
130 135 140
Ser Gly Glu Cys Cys Lys Ala Cys Asn Leu Gly Glu Gly Val Ala Gln
145 150 155 160
Pro Cys Gly Ala Asn Gln Thr Val Cys Glu Pro Cys Leu Asp Ser Val
165 170 175
Thr Phe Ser Asp Val Val Ser Ala Thr Glu Pro Cys Lys Pro Cys Thr
180 185 190
Glu Cys Val Gly Leu Gln Ser Met Ser Ala Pro Cys Val Glu Ala Asp
195 200 205
Asp Ala Val Cys Arg Cys Ala Tyr Gly Tyr Tyr Gln Asp Glu Thr Thr
210 215 220
Gly Arg Cys Glu Ala Cys Arg Val Cys Glu Ala Gly Ser Gly Leu Val
225 230 235 240
Phe Ser Cys Gln Asp Arg Gln Asn Thr Val Cys Glu Glu Cys Pro Asp
245 250 255
Gly Thr Tyr Ser Asp Glu Ala Asn His Val Asp Pro Cys Leu Pro Cys
260 265 270
Thr Val Cys Glu Asp Thr Glu Arg Gln Leu Arg Glu Cys Thr Arg Trp
275 280 285
Ala Asp Ala Glu Cys Glu Glu Arg Pro Pro Asp Cys Pro Lys Cys Pro
290 295 300
Ala Pro Glu Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys
305 310 315 320
Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val
325 330 335
Val Val Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe
340 345 350
Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu
355 360 365
Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His
370 375 380
Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Lys
385 390 395 400
Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln
405 410 415
Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu
420 425 430
Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro
435 440 445
Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu
450 455 460
Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr
465 470 475 480
Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly
485 490 495
Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr
500 505 510
Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys Ser Gly Gly Gly Ser
515 520 525
Gly Gly Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu
530 535 540
Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro
545 550 555 560
Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe
565 570 575
Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His
580 585 590
Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr
595 600 605
Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met
610 615 620
Ala Ala Phe Met Asp Asn Pro
625 630
<210> 141
<211> 631
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Canine NGFR ECD-Canines with Signal sequences
TrkA ECD v 3-variant canine IgG-B Fc-canine TrkA ECD v3
Protein A + C1q-CD16-
<400> 141
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Ala Lys Glu Ala Cys Pro Thr Gly Leu Tyr
20 25 30
Thr His Ser Gly Glu Cys Cys Lys Ala Cys Asn Leu Gly Glu Gly Val
35 40 45
Ala Gln Pro Cys Gly Ala Asn Gln Thr Val Cys Glu Pro Cys Leu Asp
50 55 60
Ser Val Thr Phe Ser Asp Val Val Ser Ala Thr Glu Pro Cys Lys Pro
65 70 75 80
Cys Thr Glu Cys Val Gly Leu Gln Ser Met Ser Ala Pro Cys Val Glu
85 90 95
Ala Asp Asp Ala Val Cys Arg Cys Ala Tyr Gly Tyr Tyr Gln Asp Glu
100 105 110
Thr Thr Gly Arg Cys Glu Ala Cys Arg Val Cys Glu Ala Gly Ser Gly
115 120 125
Leu Val Phe Ser Cys Gln Asp Arg Gln Asn Thr Val Cys Glu Glu Cys
130 135 140
Pro Asp Gly Thr Tyr Ser Asp Glu Ala Asn His Val Asp Pro Cys Leu
145 150 155 160
Pro Cys Thr Val Cys Glu Asp Thr Glu Arg Gln Leu Arg Glu Cys Thr
165 170 175
Arg Trp Ala Asp Ala Glu Cys Glu Glu Ser Gly Gly Gly Ser Gly Gly
180 185 190
Gly Ser Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu
195 200 205
Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro
210 215 220
Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe
225 230 235 240
Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His
245 250 255
Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr
260 265 270
Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met
275 280 285
Ala Ala Phe Met Asp Asn Pro Arg Pro Pro Asp Cys Pro Lys Cys Pro
290 295 300
Ala Pro Glu Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys
305 310 315 320
Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val
325 330 335
Val Val Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe
340 345 350
Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu
355 360 365
Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His
370 375 380
Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Lys
385 390 395 400
Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln
405 410 415
Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu
420 425 430
Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro
435 440 445
Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu
450 455 460
Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr
465 470 475 480
Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly
485 490 495
Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr
500 505 510
Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys Ser Gly Gly Gly Ser
515 520 525
Gly Gly Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu
530 535 540
Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro
545 550 555 560
Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe
565 570 575
Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His
580 585 590
Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr
595 600 605
Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met
610 615 620
Ala Ala Phe Met Asp Asn Pro
625 630
<210> 142
<211> 530
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine with signal sequence
IgG-B Fc-canine NGFR ECD protein A + C1q-CD16-
<400> 142
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu
130 135 140
Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
145 150 155 160
Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
165 170 175
Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly
180 185 190
Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn
195 200 205
Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp
210 215 220
Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Lys Ala Leu Pro
225 230 235 240
Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln
245 250 255
Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn
260 265 270
Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile
275 280 285
Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr
290 295 300
Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr
305 310 315 320
Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe
325 330 335
Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu
340 345 350
Ser Leu Ser His Ser Pro Gly Lys Ser Gly Gly Gly Ser Gly Gly Ala
355 360 365
Lys Glu Ala Cys Pro Thr Gly Leu Tyr Thr His Ser Gly Glu Cys Cys
370 375 380
Lys Ala Cys Asn Leu Gly Glu Gly Val Ala Gln Pro Cys Gly Ala Asn
385 390 395 400
Gln Thr Val Cys Glu Pro Cys Leu Asp Ser Val Thr Phe Ser Asp Val
405 410 415
Val Ser Ala Thr Glu Pro Cys Lys Pro Cys Thr Glu Cys Val Gly Leu
420 425 430
Gln Ser Met Ser Ala Pro Cys Val Glu Ala Asp Asp Ala Val Cys Arg
435 440 445
Cys Ala Tyr Gly Tyr Tyr Gln Asp Glu Thr Thr Gly Arg Cys Glu Ala
450 455 460
Cys Arg Val Cys Glu Ala Gly Ser Gly Leu Val Phe Ser Cys Gln Asp
465 470 475 480
Arg Gln Asn Thr Val Cys Glu Glu Cys Pro Asp Gly Thr Tyr Ser Asp
485 490 495
Glu Ala Asn His Val Asp Pro Cys Leu Pro Cys Thr Val Cys Glu Asp
500 505 510
Thr Glu Arg Gln Leu Arg Glu Cys Thr Arg Trp Ala Asp Ala Glu Cys
515 520 525
Glu Glu
530
<210> 143
<211> 4
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 143
Gly Ser Gly Ser
1
<210> 144
<211> 6
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 144
Gly Ser Gly Ser Gly Ser
1 5
<210> 145
<211> 6
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 145
Gly Gly Ser Gly Gly Ser
1 5
<210> 146
<211> 9
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 146
Gly Gly Ser Gly Gly Ser Gly Gly Ser
1 5
<210> 147
<211> 4
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 147
Gly Gly Gly Ser
1
<210> 148
<211> 8
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 148
Gly Gly Gly Ser Gly Gly Gly Ser
1 5
<210> 149
<211> 12
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 149
Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser
1 5 10
<210> 150
<211> 6
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 150
Gly Ser Ser Gly Ser Ser
1 5
<210> 151
<211> 9
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 151
Gly Ser Ser Gly Ser Ser Gly Ser Ser
1 5
<210> 152
<211> 4
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 152
Gly Gly Ser Ser
1
<210> 153
<211> 8
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 153
Gly Gly Ser Ser Gly Gly Ser Ser
1 5
<210> 154
<211> 11
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 154
Gly Gly Ser Ser Gly Gly Ser Ser Gly Gly Ser
1 5 10
<210> 155
<211> 7
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 155
Ser Gly Gly Ser Gly Gly Ser
1 5
<210> 156
<211> 9
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 156
Ser Gly Gly Gly Ser Gly Gly Gly Ser
1 5
<210> 157
<211> 12
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 157
Gly Gly Ser Ser Gly Gly Ser Ser Gly Gly Ser Ser
1 5 10
<210> 158
<211> 4
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic: exemplary linker
<400> 158
Ser Gly Gly Gly
1
<210> 159
<211> 362
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-wild type canine IgG-B
Fc protein A + C1q + CD16+
<400> 159
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp Ser Gly Gly
115 120 125
Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala
130 135 140
Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro
145 150 155 160
Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val
165 170 175
Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val
180 185 190
Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln
195 200 205
Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln
210 215 220
Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala
225 230 235 240
Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala
245 250 255
His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser
260 265 270
Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro
275 280 285
Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser
290 295 300
Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe
305 310 315 320
Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp
325 330 335
Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr
340 345 350
Gln Glu Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 160
<211> 338
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-wild type canine IgG-B
Fc protein A + C1q + CD16+
<400> 160
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro
100 105 110
Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser
115 120 125
Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg
130 135 140
Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro
145 150 155 160
Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala
165 170 175
Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val
180 185 190
Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe
195 200 205
Thr Cys Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr
210 215 220
Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu
225 230 235 240
Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys
245 250 255
Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser
260 265 270
Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln
275 280 285
Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp
290 295 300
Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His
305 310 315 320
Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro
325 330 335
Gly Lys
<210> 161
<211> 336
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-wild type canine IgG-B
Fc protein A + C1q + CD16+
<400> 161
Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
100 105 110
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
115 120 125
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
130 135 140
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
145 150 155 160
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
165 170 175
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
180 185 190
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
195 200 205
Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
210 215 220
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
225 230 235 240
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
245 250 255
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
260 265 270
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
275 280 285
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
290 295 300
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
305 310 315 320
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
325 330 335
<210> 162
<211> 371
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Cat class TrkA ECD v 2-wild type Cat class IgG2Fc
Protein A + C1q-CD16-
<400> 162
Val Ser Phe Pro Ala Ser Val Gln Leu His Ala Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Ala Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Ser Val Leu Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Ser Asn Ser Thr Ser Gly Asp Ser Gly Gly
115 120 125
Gly Ser Gly Gly Gly Ser Pro Lys Thr Ala Ser Thr Ile Glu Ser Lys
130 135 140
Thr Gly Glu Gly Pro Lys Cys Pro Val Pro Glu Ile Pro Gly Ala Pro
145 150 155 160
Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Ser Ile Ser
165 170 175
Arg Thr Pro Glu Val Thr Cys Leu Val Val Asp Leu Gly Pro Asp Asp
180 185 190
Ser Asn Val Gln Ile Thr Trp Phe Val Asp Asn Thr Glu Met His Thr
195 200 205
Ala Lys Thr Arg Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val
210 215 220
Val Ser Val Leu Pro Ile Leu His Gln Asp Trp Leu Lys Gly Lys Glu
225 230 235 240
Phe Lys Cys Lys Val Asn Ser Lys Ser Leu Pro Ser Ala Met Glu Arg
245 250 255
Thr Ile Ser Lys Ala Lys Gly Gln Pro His Glu Pro Gln Val Tyr Val
260 265 270
Leu Pro Pro Thr Gln Glu Glu Leu Ser Glu Asn Lys Val Ser Val Thr
275 280 285
Cys Leu Ile Lys Gly Phe His Pro Pro Asp Ile Ala Val Glu Trp Glu
290 295 300
Ile Thr Gly Gln Pro Glu Pro Glu Asn Asn Tyr Gln Thr Thr Pro Pro
305 310 315 320
Gln Leu Asp Ser Asp Gly Thr Tyr Phe Leu Tyr Ser Arg Leu Ser Val
325 330 335
Asp Arg Ser His Trp Gln Arg Gly Asn Thr Tyr Thr Cys Ser Val Ser
340 345 350
His Glu Ala Leu His Ser His His Thr Gln Lys Ser Leu Thr Gln Ser
355 360 365
Pro Gly Lys
370
<210> 163
<211> 347
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Cat class TrkA ECD v 3-wild type Cat class IgG2Fc
Protein A + C1q-CD16-
<400> 163
Val Ser Phe Pro Ala Ser Val Gln Leu His Ala Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Ala Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Ser Val Leu Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Pro Lys
100 105 110
Thr Ala Ser Thr Ile Glu Ser Lys Thr Gly Glu Gly Pro Lys Cys Pro
115 120 125
Val Pro Glu Ile Pro Gly Ala Pro Ser Val Phe Ile Phe Pro Pro Lys
130 135 140
Pro Lys Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr Cys Leu
145 150 155 160
Val Val Asp Leu Gly Pro Asp Asp Ser Asn Val Gln Ile Thr Trp Phe
165 170 175
Val Asp Asn Thr Glu Met His Thr Ala Lys Thr Arg Pro Arg Glu Glu
180 185 190
Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile Leu His
195 200 205
Gln Asp Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn Ser Lys
210 215 220
Ser Leu Pro Ser Ala Met Glu Arg Thr Ile Ser Lys Ala Lys Gly Gln
225 230 235 240
Pro His Glu Pro Gln Val Tyr Val Leu Pro Pro Thr Gln Glu Glu Leu
245 250 255
Ser Glu Asn Lys Val Ser Val Thr Cys Leu Ile Lys Gly Phe His Pro
260 265 270
Pro Asp Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu Pro Glu
275 280 285
Asn Asn Tyr Gln Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly Thr Tyr
290 295 300
Phe Leu Tyr Ser Arg Leu Ser Val Asp Arg Ser His Trp Gln Arg Gly
305 310 315 320
Asn Thr Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser His His
325 330 335
Thr Gln Lys Ser Leu Thr Gln Ser Pro Gly Lys
340 345
<210> 164
<211> 345
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Cat class TrkA ECD v 4-wild type Cat class IgG2Fc
Protein A + C1q-CD16-
<400> 164
Phe Pro Ala Ser Val Gln Leu His Ala Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Ala Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Ser Val Leu Ala Ala Phe Met
85 90 95
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Pro Lys Thr Ala
100 105 110
Ser Thr Ile Glu Ser Lys Thr Gly Glu Gly Pro Lys Cys Pro Val Pro
115 120 125
Glu Ile Pro Gly Ala Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys
130 135 140
Asp Thr Leu Ser Ile Ser Arg Thr Pro Glu Val Thr Cys Leu Val Val
145 150 155 160
Asp Leu Gly Pro Asp Asp Ser Asn Val Gln Ile Thr Trp Phe Val Asp
165 170 175
Asn Thr Glu Met His Thr Ala Lys Thr Arg Pro Arg Glu Glu Gln Phe
180 185 190
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile Leu His Gln Asp
195 200 205
Trp Leu Lys Gly Lys Glu Phe Lys Cys Lys Val Asn Ser Lys Ser Leu
210 215 220
Pro Ser Ala Met Glu Arg Thr Ile Ser Lys Ala Lys Gly Gln Pro His
225 230 235 240
Glu Pro Gln Val Tyr Val Leu Pro Pro Thr Gln Glu Glu Leu Ser Glu
245 250 255
Asn Lys Val Ser Val Thr Cys Leu Ile Lys Gly Phe His Pro Pro Asp
260 265 270
Ile Ala Val Glu Trp Glu Ile Thr Gly Gln Pro Glu Pro Glu Asn Asn
275 280 285
Tyr Gln Thr Thr Pro Pro Gln Leu Asp Ser Asp Gly Thr Tyr Phe Leu
290 295 300
Tyr Ser Arg Leu Ser Val Asp Arg Ser His Trp Gln Arg Gly Asn Thr
305 310 315 320
Tyr Thr Cys Ser Val Ser His Glu Ala Leu His Ser His His Thr Gln
325 330 335
Lys Ser Leu Thr Gln Ser Pro Gly Lys
340 345
<210> 165
<211> 369
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD v 2-variant equine IgG2Fc
Protein A + C1q-CD16-
<400> 165
Val Ser Phe Pro Ala Ser Val His Leu Gln Thr Ala Val Glu Gln His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Thr Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Ser Ala Ala Asn Glu Thr Met Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser Val Met Val Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Arg Asp Pro Pro Cys
115 120 125
Val Leu Ser Ala Glu Gly Val Ile Pro Ile Pro Ser Val Pro Lys Pro
130 135 140
Gln Cys Pro Pro Tyr Thr His Ser Lys Phe Leu Gly Gly Pro Ser Val
145 150 155 160
Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
165 170 175
Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln Tyr Pro Asp
180 185 190
Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His Ser Ala Ile
195 200 205
Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg Val Val Ser
210 215 220
Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys Glu Phe Lys
225 230 235 240
Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser Arg Ala Ile
245 250 255
Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr Val Leu Pro
260 265 270
Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val Thr Cys Leu
275 280 285
Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp Gln Ser Asn
290 295 300
Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro Ala Gln Leu
305 310 315 320
Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Leu Glu Thr
325 330 335
Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val Met His Glu
340 345 350
Ala Leu His Asn His Tyr Thr Lys Thr Asp Ile Ser Glu Ser Leu Gly
355 360 365
Lys
<210> 166
<211> 345
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD v 3-variant equine IgG2Fc
Protein A + C1q-CD16-
<400> 166
Val Ser Phe Pro Ala Ser Val His Leu Gln Thr Ala Val Glu Gln His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Thr Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Ser Ala Ala Asn Glu Thr Met Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser Val Met Val Ala
85 90 95
Phe Met Asp Asn Pro Pro Pro Cys Val Leu Ser Ala Glu Gly Val Ile
100 105 110
Pro Ile Pro Ser Val Pro Lys Pro Gln Cys Pro Pro Tyr Thr His Ser
115 120 125
Lys Phe Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys
130 135 140
Asp Thr Leu Met Ile Ser Arg Thr Pro Val Val Thr Cys Val Val Val
145 150 155 160
Asn Leu Ser Asp Gln Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp
165 170 175
Asn Thr Glu Val His Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe
180 185 190
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gln His Gln Asp
195 200 205
Trp Leu Ser Gly Lys Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val
210 215 220
Pro Gln Pro Ile Ser Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg
225 230 235 240
Val Pro Gln Val Tyr Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys
245 250 255
Ser Lys Val Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp
260 265 270
Ile Ser Val Glu Trp Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys
275 280 285
Tyr Ser Thr Thr Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu
290 295 300
Tyr Ser Lys Leu Ser Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser
305 310 315 320
Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Lys
325 330 335
Thr Asp Ile Ser Glu Ser Leu Gly Lys
340 345
<210> 167
<211> 343
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD v 4-variant equine IgG2Fc
Protein A + C1q-CD16-
<400> 167
Phe Pro Ala Ser Val His Leu Gln Thr Ala Val Glu Gln His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Thr Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Ser Ala Ala Asn Glu Thr Met Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser Val Met Val Ala Phe Met
85 90 95
Asp Asn Pro Pro Pro Cys Val Leu Ser Ala Glu Gly Val Ile Pro Ile
100 105 110
Pro Ser Val Pro Lys Pro Gln Cys Pro Pro Tyr Thr His Ser Lys Phe
115 120 125
Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr
130 135 140
Leu Met Ile Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu
145 150 155 160
Ser Asp Gln Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr
165 170 175
Glu Val His Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser
180 185 190
Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu
195 200 205
Ser Gly Lys Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln
210 215 220
Pro Ile Ser Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro
225 230 235 240
Gln Val Tyr Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys
245 250 255
Val Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser
260 265 270
Val Glu Trp Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser
275 280 285
Thr Thr Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser
290 295 300
Lys Leu Ser Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr
305 310 315 320
Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Lys Thr Asp
325 330 335
Ile Ser Glu Ser Leu Gly Lys
340
<210> 168
<211> 362
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine IgG-B Fc
Protein A + C1q-CD16-
<400> 168
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp Ser Gly Gly
115 120 125
Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala
130 135 140
Pro Glu Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro
145 150 155 160
Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val
165 170 175
Val Asp Leu Asp Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val
180 185 190
Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln
195 200 205
Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln
210 215 220
Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Lys Ala
225 230 235 240
Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala
245 250 255
His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser
260 265 270
Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro
275 280 285
Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser
290 295 300
Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe
305 310 315 320
Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp
325 330 335
Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr
340 345 350
Gln Glu Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 169
<211> 338
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-B Fc
Protein A + C1q-CD16-
<400> 169
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro
100 105 110
Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser
115 120 125
Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg
130 135 140
Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp Arg Glu Asp Pro
145 150 155 160
Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala
165 170 175
Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val
180 185 190
Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe
195 200 205
Thr Cys Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr
210 215 220
Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu
225 230 235 240
Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys
245 250 255
Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser
260 265 270
Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln
275 280 285
Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp
290 295 300
Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His
305 310 315 320
Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro
325 330 335
Gly Lys
<210> 170
<211> 336
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine IgG-B Fc
Protein A + C1q-CD16-
<400> 170
Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
100 105 110
Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser Val Phe
115 120 125
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
130 135 140
Glu Val Thr Cys Val Val Val Asp Leu Asp Arg Glu Asp Pro Glu Val
145 150 155 160
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
165 170 175
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
180 185 190
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
195 200 205
Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
210 215 220
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
225 230 235 240
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
245 250 255
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
260 265 270
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
275 280 285
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
290 295 300
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
305 310 315 320
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
325 330 335
<210> 171
<211> 366
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine IgG-A Fc
C1 q-protein A +
<400> 171
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp Ser Gly Gly
115 120 125
Gly Ser Gly Gly Gly Ser Phe Asn Glu Cys Arg Cys Thr Asp Thr Pro
130 135 140
Cys Pro Val Pro Glu Pro Leu Gly Gly Pro Ser Val Leu Ile Phe Pro
145 150 155 160
Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr
165 170 175
Cys Val Val Leu Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser
180 185 190
Trp Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Ser Arg
195 200 205
Glu Gln Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile
210 215 220
Gly His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val Asn
225 230 235 240
His Ile Asp Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg
245 250 255
Gly Arg Ala His Lys Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys
260 265 270
Glu Leu Ser Ser Ser Asp Thr Val Ser Ile Thr Cys Leu Ile Lys Asp
275 280 285
Phe Tyr Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln
290 295 300
Glu Pro Glu Arg Lys His Arg Met Thr Pro Pro Gln Leu Asp Glu Asp
305 310 315 320
Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp
325 330 335
Gln Gln Gly Asp Pro Phe Thr Cys Ala Val Met His Glu Ala Leu His
340 345 350
Asn His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
355 360 365
<210> 172
<211> 342
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-A Fc
C1 q-protein A +
<400> 172
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Phe Asn
100 105 110
Glu Cys Arg Cys Thr Asp Thr Pro Cys Pro Val Pro Glu Pro Leu Gly
115 120 125
Gly Pro Ser Val Leu Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu
130 135 140
Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Leu Asp Leu Gly Arg
145 150 155 160
Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Glu Val
165 170 175
His Thr Ala Lys Thr Gln Ser Arg Glu Gln Gln Phe Asn Gly Thr Tyr
180 185 190
Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Thr Gly
195 200 205
Lys Glu Phe Lys Cys Arg Val Asn His Ile Asp Leu Pro Ser Pro Ile
210 215 220
Glu Arg Thr Ile Ser Lys Ala Arg Gly Arg Ala His Lys Pro Ser Val
225 230 235 240
Tyr Val Leu Pro Pro Ser Pro Lys Glu Leu Ser Ser Ser Asp Thr Val
245 250 255
Ser Ile Thr Cys Leu Ile Lys Asp Phe Tyr Pro Pro Asp Ile Asp Val
260 265 270
Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Arg Lys His Arg Met
275 280 285
Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
290 295 300
Leu Ser Val Asp Lys Ser Arg Trp Gln Gln Gly Asp Pro Phe Thr Cys
305 310 315 320
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Asp Leu Ser Leu
325 330 335
Ser His Ser Pro Gly Lys
340
<210> 173
<211> 340
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine IgG-A Fc
C1 q-protein A +
<400> 173
Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Phe Asn Glu Cys
100 105 110
Arg Cys Thr Asp Thr Pro Cys Pro Val Pro Glu Pro Leu Gly Gly Pro
115 120 125
Ser Val Leu Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala
130 135 140
Arg Thr Pro Glu Val Thr Cys Val Val Leu Asp Leu Gly Arg Glu Asp
145 150 155 160
Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Glu Val His Thr
165 170 175
Ala Lys Thr Gln Ser Arg Glu Gln Gln Phe Asn Gly Thr Tyr Arg Val
180 185 190
Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Thr Gly Lys Glu
195 200 205
Phe Lys Cys Arg Val Asn His Ile Asp Leu Pro Ser Pro Ile Glu Arg
210 215 220
Thr Ile Ser Lys Ala Arg Gly Arg Ala His Lys Pro Ser Val Tyr Val
225 230 235 240
Leu Pro Pro Ser Pro Lys Glu Leu Ser Ser Ser Asp Thr Val Ser Ile
245 250 255
Thr Cys Leu Ile Lys Asp Phe Tyr Pro Pro Asp Ile Asp Val Glu Trp
260 265 270
Gln Ser Asn Gly Gln Gln Glu Pro Glu Arg Lys His Arg Met Thr Pro
275 280 285
Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser
290 295 300
Val Asp Lys Ser Arg Trp Gln Gln Gly Asp Pro Phe Thr Cys Ala Val
305 310 315 320
Met His Glu Ala Leu His Asn His Tyr Thr Asp Leu Ser Leu Ser His
325 330 335
Ser Pro Gly Lys
340
<210> 174
<211> 367
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine IgG-D Fc
C1 q-protein A +
<400> 174
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp Ser Gly Gly
115 120 125
Gly Ser Gly Gly Gly Ser Pro Lys Glu Ser Thr Cys Lys Cys Ile Ser
130 135 140
Pro Cys Pro Val Pro Glu Ser Leu Gly Gly Pro Ser Val Phe Ile Phe
145 150 155 160
Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Ile
165 170 175
Thr Cys Val Val Leu Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile
180 185 190
Ser Trp Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Pro
195 200 205
Arg Glu Gln Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro
210 215 220
Ile Gly His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val
225 230 235 240
Asn His Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala
245 250 255
Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Pro
260 265 270
Lys Glu Leu Ser Ser Ser Asp Thr Val Thr Leu Thr Cys Leu Ile Lys
275 280 285
Asp Phe Phe Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln
290 295 300
Pro Glu Pro Glu Ser Lys Tyr His Thr Thr Ala Pro Gln Leu Asp Glu
305 310 315 320
Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg
325 330 335
Trp Gln Gln Gly Asp Thr Phe Thr Cys Ala Val Met His Glu Ala Leu
340 345 350
His Asn His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
355 360 365
<210> 175
<211> 343
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-D Fc
C1 q-protein A +
<400> 175
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Pro Lys
100 105 110
Glu Ser Thr Cys Lys Cys Ile Ser Pro Cys Pro Val Pro Glu Ser Leu
115 120 125
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu
130 135 140
Leu Ile Ala Arg Thr Pro Glu Ile Thr Cys Val Val Leu Asp Leu Gly
145 150 155 160
Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Glu
165 170 175
Val His Thr Ala Lys Thr Gln Pro Arg Glu Gln Gln Phe Asn Ser Thr
180 185 190
Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Thr
195 200 205
Gly Lys Glu Phe Lys Cys Arg Val Asn His Ile Gly Leu Pro Ser Pro
210 215 220
Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser
225 230 235 240
Val Tyr Val Leu Pro Pro Ser Pro Lys Glu Leu Ser Ser Ser Asp Thr
245 250 255
Val Thr Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Glu Ile Asp
260 265 270
Val Glu Trp Gln Ser Asn Gly Gln Pro Glu Pro Glu Ser Lys Tyr His
275 280 285
Thr Thr Ala Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser
290 295 300
Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Gln Gly Asp Thr Phe Thr
305 310 315 320
Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Asp Leu Ser
325 330 335
Leu Ser His Ser Pro Gly Lys
340
<210> 176
<211> 341
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine IgG-D Fc
C1 q-protein A +
<400> 176
Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Pro Lys Glu Ser
100 105 110
Thr Cys Lys Cys Ile Ser Pro Cys Pro Val Pro Glu Ser Leu Gly Gly
115 120 125
Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile
130 135 140
Ala Arg Thr Pro Glu Ile Thr Cys Val Val Leu Asp Leu Gly Arg Glu
145 150 155 160
Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Glu Val His
165 170 175
Thr Ala Lys Thr Gln Pro Arg Glu Gln Gln Phe Asn Ser Thr Tyr Arg
180 185 190
Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Thr Gly Lys
195 200 205
Glu Phe Lys Cys Arg Val Asn His Ile Gly Leu Pro Ser Pro Ile Glu
210 215 220
Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr
225 230 235 240
Val Leu Pro Pro Ser Pro Lys Glu Leu Ser Ser Ser Asp Thr Val Thr
245 250 255
Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Glu Ile Asp Val Glu
260 265 270
Trp Gln Ser Asn Gly Gln Pro Glu Pro Glu Ser Lys Tyr His Thr Thr
275 280 285
Ala Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu
290 295 300
Ser Val Asp Lys Ser Arg Trp Gln Gln Gly Asp Thr Phe Thr Cys Ala
305 310 315 320
Val Met His Glu Ala Leu His Asn His Tyr Thr Asp Leu Ser Leu Ser
325 330 335
His Ser Pro Gly Lys
340
<210> 177
<211> 353
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD v 2-variant with IgG1 hinge
Horse class IgG 2C 1 q-protein A +
<400> 177
Val Ser Phe Pro Ala Ser Val His Leu Gln Thr Ala Val Glu Gln His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Thr Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Ser Ala Ala Asn Glu Thr Met Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser Val Met Val Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Arg Asp Asp Met Ser
115 120 125
Lys Cys Pro Lys Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val
130 135 140
Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
145 150 155 160
Pro Val Val Thr Cys Val Val Val Asn Leu Ser Asp Gln Tyr Pro Asp
165 170 175
Val Gln Phe Ser Trp Tyr Val Asp Asn Thr Glu Val His Ser Ala Ile
180 185 190
Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser Thr Tyr Arg Val Val Ser
195 200 205
Val Leu Pro Ile Gln His Gln Asp Trp Leu Ser Gly Lys Glu Phe Lys
210 215 220
Cys Ser Val Thr Asn Val Gly Val Pro Gln Pro Ile Ser Arg Ala Ile
225 230 235 240
Ser Arg Gly Lys Gly Pro Ser Arg Val Pro Gln Val Tyr Val Leu Pro
245 250 255
Pro His Pro Asp Glu Leu Ala Lys Ser Lys Val Ser Val Thr Cys Leu
260 265 270
Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser Val Glu Trp Gln Ser Asn
275 280 285
Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser Thr Thr Pro Ala Gln Leu
290 295 300
Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Leu Glu Thr
305 310 315 320
Ser Arg Trp Gln Gln Val Glu Ser Phe Thr Cys Ala Val Met His Glu
325 330 335
Ala Leu His Asn His Tyr Thr Lys Thr Asp Ile Ser Glu Ser Leu Gly
340 345 350
Lys
<210> 178
<211> 329
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD v 3-variant with IgG1 hinge
Horse class IgG 2C 1 q-protein A +
<400> 178
Val Ser Phe Pro Ala Ser Val His Leu Gln Thr Ala Val Glu Gln His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Thr Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Ser Ala Ala Asn Glu Thr Met Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser Val Met Val Ala
85 90 95
Phe Met Asp Asn Pro Asp Met Ser Lys Cys Pro Lys Cys Pro Ala Pro
100 105 110
Glu Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys
115 120 125
Asp Thr Leu Met Ile Ser Arg Thr Pro Val Val Thr Cys Val Val Val
130 135 140
Asn Leu Ser Asp Gln Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp
145 150 155 160
Asn Thr Glu Val His Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe
165 170 175
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gln His Gln Asp
180 185 190
Trp Leu Ser Gly Lys Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val
195 200 205
Pro Gln Pro Ile Ser Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg
210 215 220
Val Pro Gln Val Tyr Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys
225 230 235 240
Ser Lys Val Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp
245 250 255
Ile Ser Val Glu Trp Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys
260 265 270
Tyr Ser Thr Thr Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu
275 280 285
Tyr Ser Lys Leu Ser Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser
290 295 300
Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Lys
305 310 315 320
Thr Asp Ile Ser Glu Ser Leu Gly Lys
325
<210> 179
<211> 327
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary equine TrkA ECD v 4-variant equine with IgG1 hinge
IgG 2C 1 q-protein A +
<400> 179
Phe Pro Ala Ser Val His Leu Gln Thr Ala Val Glu Gln His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Thr Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Ser Ala Ala Asn Glu Thr Met Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Thr Asn Pro Tyr Gly Gln Asp Ser Ala Ser Val Met Val Ala Phe Met
85 90 95
Asp Asn Pro Asp Met Ser Lys Cys Pro Lys Cys Pro Ala Pro Glu Leu
100 105 110
Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Asn Pro Lys Asp Thr
115 120 125
Leu Met Ile Ser Arg Thr Pro Val Val Thr Cys Val Val Val Asn Leu
130 135 140
Ser Asp Gln Tyr Pro Asp Val Gln Phe Ser Trp Tyr Val Asp Asn Thr
145 150 155 160
Glu Val His Ser Ala Ile Thr Lys Gln Arg Glu Ala Gln Phe Asn Ser
165 170 175
Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gln His Gln Asp Trp Leu
180 185 190
Ser Gly Lys Glu Phe Lys Cys Ser Val Thr Asn Val Gly Val Pro Gln
195 200 205
Pro Ile Ser Arg Ala Ile Ser Arg Gly Lys Gly Pro Ser Arg Val Pro
210 215 220
Gln Val Tyr Val Leu Pro Pro His Pro Asp Glu Leu Ala Lys Ser Lys
225 230 235 240
Val Ser Val Thr Cys Leu Val Lys Asp Phe Tyr Pro Pro Asp Ile Ser
245 250 255
Val Glu Trp Gln Ser Asn Arg Trp Pro Glu Leu Glu Gly Lys Tyr Ser
260 265 270
Thr Thr Pro Ala Gln Leu Asp Gly Asp Gly Ser Tyr Phe Leu Tyr Ser
275 280 285
Lys Leu Ser Leu Glu Thr Ser Arg Trp Gln Gln Val Glu Ser Phe Thr
290 295 300
Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Lys Thr Asp
305 310 315 320
Ile Ser Glu Ser Leu Gly Lys
325
<210> 180
<211> 362
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine IgG-B Fc
Additional disulfide protein A + C1q-CD16-
<400> 180
Val Ser Phe Pro Ala Ser Cys Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Cys Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp Ser Gly Gly
115 120 125
Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala
130 135 140
Pro Glu Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro
145 150 155 160
Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val
165 170 175
Val Asp Leu Asp Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val
180 185 190
Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln
195 200 205
Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln
210 215 220
Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Lys Ala
225 230 235 240
Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala
245 250 255
His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser
260 265 270
Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro
275 280 285
Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser
290 295 300
Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe
305 310 315 320
Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp
325 330 335
Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr
340 345 350
Gln Glu Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 181
<211> 338
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-B Fc
Additional disulfide protein A + C1q-CD16-
<400> 181
Val Ser Phe Pro Ala Ser Cys Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Cys Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro
100 105 110
Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser
115 120 125
Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg
130 135 140
Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp Arg Glu Asp Pro
145 150 155 160
Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala
165 170 175
Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val
180 185 190
Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe
195 200 205
Thr Cys Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr
210 215 220
Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu
225 230 235 240
Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys
245 250 255
Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser
260 265 270
Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln
275 280 285
Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp
290 295 300
Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His
305 310 315 320
Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro
325 330 335
Gly Lys
<210> 182
<211> 336
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine IgG-B Fc
Additional disulfide protein A + C1q-CD16-
<400> 182
Phe Pro Ala Ser Cys Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Cys Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
100 105 110
Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser Val Phe
115 120 125
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
130 135 140
Glu Val Thr Cys Val Val Val Asp Leu Asp Arg Glu Asp Pro Glu Val
145 150 155 160
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
165 170 175
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
180 185 190
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
195 200 205
Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
210 215 220
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
225 230 235 240
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
245 250 255
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
260 265 270
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
275 280 285
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
290 295 300
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
305 310 315 320
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
325 330 335
<210> 183
<211> 362
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine IgG-B Fc
Protein A + C1q-CD16-
<400> 183
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp Ser Gly Gly
115 120 125
Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala
130 135 140
Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro
145 150 155 160
Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val
165 170 175
Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val
180 185 190
Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln
195 200 205
Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln
210 215 220
Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Ile Gly
225 230 235 240
Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala
245 250 255
His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser
260 265 270
Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro
275 280 285
Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser
290 295 300
Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe
305 310 315 320
Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp
325 330 335
Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr
340 345 350
Gln Glu Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 184
<211> 338
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-B Fc
Protein A + C1q-CD16-
<400> 184
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro
100 105 110
Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser
115 120 125
Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg
130 135 140
Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro
145 150 155 160
Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala
165 170 175
Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val
180 185 190
Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe
195 200 205
Thr Cys Arg Val Asn Asn Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr
210 215 220
Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu
225 230 235 240
Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys
245 250 255
Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser
260 265 270
Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln
275 280 285
Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp
290 295 300
Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His
305 310 315 320
Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro
325 330 335
Gly Lys
<210> 185
<211> 336
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine IgG-B Fc
Protein A + C1q-CD16-
<400> 185
Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
100 105 110
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
115 120 125
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
130 135 140
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
145 150 155 160
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
165 170 175
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
180 185 190
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
195 200 205
Arg Val Asn Asn Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
210 215 220
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
225 230 235 240
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
245 250 255
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
260 265 270
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
275 280 285
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
290 295 300
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
305 310 315 320
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
325 330 335
<210> 186
<211> 362
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine IgG-B Fc
Protein A + C1q-CD16-
<400> 186
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp Ser Gly Gly
115 120 125
Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala
130 135 140
Pro Glu Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro
145 150 155 160
Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val
165 170 175
Val Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val
180 185 190
Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln
195 200 205
Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln
210 215 220
Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Lys Ala
225 230 235 240
Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala
245 250 255
His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser
260 265 270
Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro
275 280 285
Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser
290 295 300
Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe
305 310 315 320
Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp
325 330 335
Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr
340 345 350
Gln Glu Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 187
<211> 338
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-B Fc
Protein A + C1q-CD16-
<400> 187
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro
100 105 110
Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser
115 120 125
Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg
130 135 140
Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Gly Arg Glu Asp Pro
145 150 155 160
Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala
165 170 175
Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val
180 185 190
Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe
195 200 205
Thr Cys Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr
210 215 220
Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu
225 230 235 240
Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys
245 250 255
Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser
260 265 270
Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln
275 280 285
Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp
290 295 300
Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His
305 310 315 320
Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro
325 330 335
Gly Lys
<210> 188
<211> 336
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine IgG-B Fc
Protein A + C1q-CD16-
<400> 188
Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
100 105 110
Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser Val Phe
115 120 125
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
130 135 140
Glu Val Thr Cys Val Val Val Asp Leu Gly Arg Glu Asp Pro Glu Val
145 150 155 160
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
165 170 175
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
180 185 190
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
195 200 205
Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
210 215 220
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
225 230 235 240
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
245 250 255
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
260 265 270
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
275 280 285
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
290 295 300
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
305 310 315 320
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
325 330 335
<210> 189
<211> 448
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3x 2-variant canine IgG-B
Fc protein A + C1q-CD16-
<400> 189
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Val Ser
100 105 110
Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp
115 120 125
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
130 135 140
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
145 150 155 160
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
165 170 175
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
180 185 190
Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met
195 200 205
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
210 215 220
Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Leu Gly Arg Glu Asp Pro Glu Val
260 265 270
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
275 280 285
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
305 310 315 320
Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
325 330 335
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
340 345 350
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
355 360 365
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
370 375 380
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
385 390 395 400
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
405 410 415
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
420 425 430
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
435 440 445
<210> 190
<211> 345
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variants Canine IgG-B Fc-Canine TrkA ECD v3
Protein A + C1q-CD16-
<400> 190
Pro Lys Arg Glu Asn Gly Arg Val Pro Arg Pro Pro Asp Cys Pro Lys
1 5 10 15
Cys Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser Val Phe Ile Phe Pro
20 25 30
Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr
35 40 45
Cys Val Val Val Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser
50 55 60
Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg
65 70 75 80
Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile
85 90 95
Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn
100 105 110
Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg
115 120 125
Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu
130 135 140
Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe
145 150 155 160
Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu
165 170 175
Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly
180 185 190
Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln
195 200 205
Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn
210 215 220
His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys Ser Gly Gly
225 230 235 240
Gly Ser Gly Gly Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala
245 250 255
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
260 265 270
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
275 280 285
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
290 295 300
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
305 310 315 320
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe
325 330 335
Val Met Ala Ala Phe Met Asp Asn Pro
340 345
<210> 191
<211> 446
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-B Fc
The canine TrkA ECD v3 protein A + C1q-CD16-
<400> 191
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro
100 105 110
Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser
115 120 125
Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg
130 135 140
Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Gly Arg Glu Asp Pro
145 150 155 160
Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala
165 170 175
Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val
180 185 190
Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe
195 200 205
Thr Cys Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr
210 215 220
Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu
225 230 235 240
Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys
245 250 255
Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser
260 265 270
Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln
275 280 285
Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp
290 295 300
Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His
305 310 315 320
Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro
325 330 335
Gly Lys Ser Gly Gly Gly Ser Gly Gly Val Ser Phe Pro Ala Ser Val
340 345 350
Gln Leu His Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser
355 360 365
Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser
370 375 380
Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val
385 390 395 400
Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr
405 410 415
His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly
420 425 430
Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro
435 440 445
<210> 192
<211> 609
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-canine NGFR ECD-
Variant canine IgG-B Fc-canine TrkA ECD v3 protein A + C1q-
CD16 -
<400> 192
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Ala Lys
100 105 110
Glu Ala Cys Pro Thr Gly Leu Tyr Thr His Ser Gly Glu Cys Cys Lys
115 120 125
Ala Cys Asn Leu Gly Glu Gly Val Ala Gln Pro Cys Gly Ala Asn Gln
130 135 140
Thr Val Cys Glu Pro Cys Leu Asp Ser Val Thr Phe Ser Asp Val Val
145 150 155 160
Ser Ala Thr Glu Pro Cys Lys Pro Cys Thr Glu Cys Val Gly Leu Gln
165 170 175
Ser Met Ser Ala Pro Cys Val Glu Ala Asp Asp Ala Val Cys Arg Cys
180 185 190
Ala Tyr Gly Tyr Tyr Gln Asp Glu Thr Thr Gly Arg Cys Glu Ala Cys
195 200 205
Arg Val Cys Glu Ala Gly Ser Gly Leu Val Phe Ser Cys Gln Asp Arg
210 215 220
Gln Asn Thr Val Cys Glu Glu Cys Pro Asp Gly Thr Tyr Ser Asp Glu
225 230 235 240
Ala Asn His Val Asp Pro Cys Leu Pro Cys Thr Val Cys Glu Asp Thr
245 250 255
Glu Arg Gln Leu Arg Glu Cys Thr Arg Trp Ala Asp Ala Glu Cys Glu
260 265 270
Glu Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly
275 280 285
Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu
290 295 300
Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Gly Arg
305 310 315 320
Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met
325 330 335
Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr
340 345 350
Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly
355 360 365
Lys Gln Phe Thr Cys Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile
370 375 380
Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val
385 390 395 400
Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser
405 410 415
Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu
420 425 430
Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr
435 440 445
Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu
450 455 460
Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala
465 470 475 480
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser
485 490 495
His Ser Pro Gly Lys Ser Gly Gly Gly Ser Gly Gly Val Ser Phe Pro
500 505 510
Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp Cys Ile
515 520 525
Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe
530 535 540
Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu
545 550 555 560
Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg Leu Asn
565 570 575
Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn
580 585 590
Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met Asp Asn
595 600 605
Pro
<210> 193
<211> 609
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine NGFR ECD-canine TrkA ECD v3
Variant canine IgG-B Fc-canine TrkA ECD v3 protein A + C1q-
CD16 -
<400> 193
Ala Lys Glu Ala Cys Pro Thr Gly Leu Tyr Thr His Ser Gly Glu Cys
1 5 10 15
Cys Lys Ala Cys Asn Leu Gly Glu Gly Val Ala Gln Pro Cys Gly Ala
20 25 30
Asn Gln Thr Val Cys Glu Pro Cys Leu Asp Ser Val Thr Phe Ser Asp
35 40 45
Val Val Ser Ala Thr Glu Pro Cys Lys Pro Cys Thr Glu Cys Val Gly
50 55 60
Leu Gln Ser Met Ser Ala Pro Cys Val Glu Ala Asp Asp Ala Val Cys
65 70 75 80
Arg Cys Ala Tyr Gly Tyr Tyr Gln Asp Glu Thr Thr Gly Arg Cys Glu
85 90 95
Ala Cys Arg Val Cys Glu Ala Gly Ser Gly Leu Val Phe Ser Cys Gln
100 105 110
Asp Arg Gln Asn Thr Val Cys Glu Glu Cys Pro Asp Gly Thr Tyr Ser
115 120 125
Asp Glu Ala Asn His Val Asp Pro Cys Leu Pro Cys Thr Val Cys Glu
130 135 140
Asp Thr Glu Arg Gln Leu Arg Glu Cys Thr Arg Trp Ala Asp Ala Glu
145 150 155 160
Cys Glu Glu Ser Gly Gly Gly Ser Gly Gly Gly Ser Val Ser Phe Pro
165 170 175
Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp Cys Ile
180 185 190
Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe
195 200 205
Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu
210 215 220
Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg Leu Asn
225 230 235 240
Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn
245 250 255
Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met Asp Asn
260 265 270
Pro Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly
275 280 285
Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu
290 295 300
Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Gly Arg
305 310 315 320
Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met
325 330 335
Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr
340 345 350
Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly
355 360 365
Lys Gln Phe Thr Cys Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile
370 375 380
Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val
385 390 395 400
Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser
405 410 415
Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu
420 425 430
Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr
435 440 445
Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu
450 455 460
Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala
465 470 475 480
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser
485 490 495
His Ser Pro Gly Lys Ser Gly Gly Gly Ser Gly Gly Val Ser Phe Pro
500 505 510
Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp Cys Ile
515 520 525
Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe
530 535 540
Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu
545 550 555 560
Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg Leu Asn
565 570 575
Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn
580 585 590
Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met Asp Asn
595 600 605
Pro
<210> 194
<211> 508
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-B Fc
Canine NGFR ECD protein A + C1q-CD16-
<400> 194
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro
100 105 110
Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Pro Leu Gly Gly Pro Ser
115 120 125
Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg
130 135 140
Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Gly Arg Glu Asp Pro
145 150 155 160
Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala
165 170 175
Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val
180 185 190
Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe
195 200 205
Thr Cys Arg Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr
210 215 220
Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu
225 230 235 240
Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys
245 250 255
Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser
260 265 270
Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln
275 280 285
Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp
290 295 300
Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His
305 310 315 320
Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro
325 330 335
Gly Lys Ser Gly Gly Gly Ser Gly Gly Ala Lys Glu Ala Cys Pro Thr
340 345 350
Gly Leu Tyr Thr His Ser Gly Glu Cys Cys Lys Ala Cys Asn Leu Gly
355 360 365
Glu Gly Val Ala Gln Pro Cys Gly Ala Asn Gln Thr Val Cys Glu Pro
370 375 380
Cys Leu Asp Ser Val Thr Phe Ser Asp Val Val Ser Ala Thr Glu Pro
385 390 395 400
Cys Lys Pro Cys Thr Glu Cys Val Gly Leu Gln Ser Met Ser Ala Pro
405 410 415
Cys Val Glu Ala Asp Asp Ala Val Cys Arg Cys Ala Tyr Gly Tyr Tyr
420 425 430
Gln Asp Glu Thr Thr Gly Arg Cys Glu Ala Cys Arg Val Cys Glu Ala
435 440 445
Gly Ser Gly Leu Val Phe Ser Cys Gln Asp Arg Gln Asn Thr Val Cys
450 455 460
Glu Glu Cys Pro Asp Gly Thr Tyr Ser Asp Glu Ala Asn His Val Asp
465 470 475 480
Pro Cys Leu Pro Cys Thr Val Cys Glu Asp Thr Glu Arg Gln Leu Arg
485 490 495
Glu Cys Thr Arg Trp Ala Asp Ala Glu Cys Glu Glu
500 505
<210> 195
<211> 305
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Canine FcRn with Poly His
<400> 195
Met Gly Val Pro Arg Pro Arg Ser Trp Gly Leu Gly Phe Leu Leu Phe
1 5 10 15
Leu Leu Pro Thr Leu Arg Ala Ala Asp Ser His Leu Ser Leu Leu Tyr
20 25 30
His Leu Thr Ala Val Ser Ala Pro Pro Pro Gly Thr Pro Ala Phe Trp
35 40 45
Ala Ser Gly Trp Leu Gly Pro Gln Gln Tyr Leu Ser Tyr Asn Asn Leu
50 55 60
Arg Ala Gln Ala Glu Pro Tyr Gly Ala Trp Val Trp Glu Asn Gln Val
65 70 75 80
Ser Trp Tyr Trp Glu Lys Glu Thr Thr Asp Leu Arg Thr Lys Glu Gly
85 90 95
Leu Phe Leu Glu Ala Leu Lys Ala Leu Gly Asp Gly Gly Pro Tyr Thr
100 105 110
Leu Gln Gly Leu Leu Gly Cys Glu Leu Gly Pro Asp Asn Thr Ser Val
115 120 125
Pro Val Ala Lys Phe Ala Leu Asn Gly Glu Asp Phe Met Thr Phe Asp
130 135 140
Pro Lys Leu Gly Thr Trp Asn Gly Asp Trp Pro Glu Thr Glu Thr Val
145 150 155 160
Ser Lys Arg Trp Met Gln Gln Ala Gly Ala Val Ser Lys Glu Arg Thr
165 170 175
Phe Leu Leu Tyr Ser Cys Pro Gln Arg Leu Leu Gly His Leu Glu Arg
180 185 190
Gly Arg Gly Asn Leu Glu Trp Lys Glu Pro Pro Ser Met Arg Leu Lys
195 200 205
Ala Arg Pro Gly Ser Pro Gly Phe Ser Val Leu Thr Cys Ser Ala Phe
210 215 220
Ser Phe Tyr Pro Pro Glu Leu Gln Leu Arg Phe Leu Arg Asn Gly Leu
225 230 235 240
Ala Ala Gly Ser Gly Glu Gly Asp Phe Gly Pro Asn Gly Asp Gly Ser
245 250 255
Phe His Ala Trp Ser Ser Leu Thr Val Lys Ser Gly Asp Glu His His
260 265 270
Tyr Arg Cys Leu Val Gln His Ala Gly Leu Pro Gln Pro Leu Thr Val
275 280 285
Glu Leu Glu Ser Pro Ala Lys Ser Ser Gly Ser His His His His His
290 295 300
His
305
<210> 196
<211> 125
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary Canine B2M
<400> 196
Met Ala Pro Arg Pro Ala Leu Ala Thr Ala Gly Phe Leu Ala Leu Leu
1 5 10 15
Leu Ile Leu Leu Ala Ala Cys Arg Leu Asp Ala Val Gln His Pro Pro
20 25 30
Lys Ile Gln Val Tyr Ser Arg His Pro Ala Glu Asn Gly Lys Pro Asn
35 40 45
Phe Leu Asn Cys Tyr Val Ser Gly Phe His Pro Pro Glu Ile Glu Ile
50 55 60
Asp Leu Leu Lys Asn Gly Lys Glu Met Lys Ala Glu Gln Thr Asp Leu
65 70 75 80
Ser Phe Ser Lys Asp Trp Thr Phe Tyr Leu Leu Val His Thr Glu Phe
85 90 95
Thr Pro Asn Glu Gln Asp Glu Phe Ser Cys Arg Val Lys His Val Thr
100 105 110
Leu Ser Glu Pro Gln Ile Val Lys Trp Asp Arg Asp Asn
115 120 125
<210> 197
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q +
CD16 + L(23)F (F00)
<400> 197
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Phe Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 198
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q +
CD16 + L(23)Y (Y00)
<400> 198
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Tyr Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 199
<211> 221
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant canine IgG-A Fc (F00; protein A +;
C1q-; CD16 -) I(21)T; R(23)F; T(25)A; E(80)G; T(205)A;
Q(207)H
<400> 199
Pro Val Pro Glu Pro Leu Gly Gly Pro Ser Val Leu Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Phe Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Leu Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Ser Arg Glu
50 55 60
Gln Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val Asn His
85 90 95
Ile Asp Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Arg Ala His Lys Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys Glu
115 120 125
Leu Ser Ser Ser Asp Thr Val Ser Ile Thr Cys Leu Ile Lys Asp Phe
130 135 140
Tyr Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu
145 150 155 160
Pro Glu Arg Lys His Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly
165 170 175
Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln
180 185 190
Gln Gly Asp Pro Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn
195 200 205
His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 200
<211> 221
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant canine IgG-A Fc (protein A +; C1q +;
CD16 +) V2A; P5M; I21T; R23L; T25A; L35V; G38D; R39P;
Q65E; E80G; R93K; H96N; I97K; D98A; T205A; Q207H
<400> 200
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Leu Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Ser Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Arg Ala His Lys Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys Glu
115 120 125
Leu Ser Ser Ser Asp Thr Val Ser Ile Thr Cys Leu Ile Lys Asp Phe
130 135 140
Tyr Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu
145 150 155 160
Pro Glu Arg Lys His Arg Met Thr Pro Pro Gln Leu Asp Glu Asp Gly
165 170 175
Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln
180 185 190
Gln Gly Asp Pro Phe Thr Cys Ala Val Met His Glu Ala Leu His Asn
195 200 205
His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 201
<211> 222
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant canine IgG-D Fc (F00; protein A +;
C1q-; CD16 -) I(21)T; R(23)F; T(25)A; E(80)G; Q(205)A;
Q(207)H
<400> 201
Cys Pro Val Pro Glu Ser Leu Gly Gly Pro Ser Val Phe Ile Phe Pro
1 5 10 15
Pro Lys Pro Lys Asp Thr Leu Phe Ile Ala Arg Thr Pro Glu Ile Thr
20 25 30
Cys Val Val Leu Asp Leu Gly Arg Glu Asp Pro Glu Val Gln Ile Ser
35 40 45
Trp Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Pro Arg
50 55 60
Glu Gln Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
65 70 75 80
Gly His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Arg Val Asn
85 90 95
His Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg
100 105 110
Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys
115 120 125
Glu Leu Ser Ser Ser Asp Thr Val Thr Leu Thr Cys Leu Ile Lys Asp
130 135 140
Phe Phe Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Pro
145 150 155 160
Glu Pro Glu Ser Lys Tyr His Thr Thr Ala Pro Gln Leu Asp Glu Asp
165 170 175
Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp
180 185 190
Gln Gln Gly Asp Thr Phe Thr Cys Ala Val Met His Glu Ala Leu His
195 200 205
Asn His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 202
<211> 222
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant canine IgG-D Fc (protein A +; C1q +;
CD16 +) V2A; S5M; I21T; R23L; T25A; L35V; G38D; R39P;
Q65E; E80G; R93K; H96N; I97K; G98A; Q207H
<400> 202
Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro
1 5 10 15
Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Ile Thr
20 25 30
Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser
35 40 45
Trp Phe Val Asp Gly Lys Glu Val His Thr Ala Lys Thr Gln Pro Arg
50 55 60
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Ile
65 70 75 80
Gly His Gln Asp Trp Leu Thr Gly Lys Glu Phe Lys Cys Lys Val Asn
85 90 95
Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg
100 105 110
Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Pro Lys
115 120 125
Glu Leu Ser Ser Ser Asp Thr Val Thr Leu Thr Cys Leu Ile Lys Asp
130 135 140
Phe Phe Pro Pro Glu Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Pro
145 150 155 160
Glu Pro Glu Ser Lys Tyr His Thr Thr Ala Pro Gln Leu Asp Glu Asp
165 170 175
Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp
180 185 190
Gln Gln Gly Asp Thr Phe Thr Cys Ala Val Met His Glu Ala Leu His
195 200 205
Asn His Tyr Thr Asp Leu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 203
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc (0Y0) protein A +
C1q + CD16 + Q(82)Y (0Y0)
<400> 203
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Tyr Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 204
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variants of canine IgG-B Fc (0YH) Gln82Tyr
Asn207His
<400> 204
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Tyr Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His His His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 205
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variants of canine IgG-B Fc (0YY) Gln82Tyr
Asn207Tyr
<400> 205
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Tyr Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Tyr His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 206
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variants Canine IgG-B Fc (00Y) Asn207Tyr
<400> 206
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Tyr His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 207
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variants Canine IgG-B Fc (YTE) Leu23Tyr
Ala25Thr Thr27Glu
<400> 207
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn
85 90 95
Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 208
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q-
CD16 - K(93)R K(97)I A(98)G L(23)F (F00)
<400> 208
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Phe Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn
85 90 95
Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 209
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q-
CD16 - K(93)R K(97)I A(98)G L(23)Y (Y00)
<400> 209
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Tyr Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn
85 90 95
Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 210
<211> 220
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary variant Canine IgG-B Fc protein A + C1q-
CD16 - K(93)R K(97)I A(98)G Q(82)Y (0Y0)
<400> 210
Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp
35 40 45
Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu
50 55 60
Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly
65 70 75 80
His Tyr Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn
85 90 95
Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly
100 105 110
Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu
115 120 125
Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe
130 135 140
Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro
145 150 155 160
Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser
165 170 175
Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg
180 185 190
Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His
195 200 205
Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
210 215 220
<210> 211
<211> 384
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine with signal sequence
IgG-B Fc protein A + C1q + CD16+ L (23) F (F00)
<400> 211
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro
115 120 125
Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr
130 135 140
Ser Gly Asp Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
145 150 155 160
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
165 170 175
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Phe Ile Ala Arg Thr Pro
180 185 190
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
195 200 205
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
210 215 220
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
225 230 235 240
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
245 250 255
Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
260 265 270
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
275 280 285
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
290 295 300
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
305 310 315 320
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
325 330 335
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
340 345 350
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
355 360 365
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
370 375 380
<210> 212
<211> 362
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine IgG-B Fc
Protein A + C1q + CD16+ L (23) F (F00)
<400> 212
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp Ser Gly Gly
115 120 125
Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala
130 135 140
Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro
145 150 155 160
Lys Asp Thr Leu Phe Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val
165 170 175
Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val
180 185 190
Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln
195 200 205
Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln
210 215 220
Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala
225 230 235 240
Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala
245 250 255
His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser
260 265 270
Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro
275 280 285
Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser
290 295 300
Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe
305 310 315 320
Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp
325 330 335
Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr
340 345 350
Gln Glu Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 213
<211> 360
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine with signal sequence
IgG-B Fc protein A + C1q + CD16+ L (23) F (F00)
<400> 213
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu
130 135 140
Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
145 150 155 160
Thr Leu Phe Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
165 170 175
Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly
180 185 190
Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn
195 200 205
Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp
210 215 220
Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala Leu Pro
225 230 235 240
Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln
245 250 255
Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn
260 265 270
Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile
275 280 285
Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr
290 295 300
Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr
305 310 315 320
Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe
325 330 335
Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu
340 345 350
Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 214
<211> 338
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-B Fc
Protein A + C1q + CD16+ L (23) F (F00)
<400> 214
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro
100 105 110
Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser
115 120 125
Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Phe Ile Ala Arg
130 135 140
Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro
145 150 155 160
Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala
165 170 175
Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val
180 185 190
Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe
195 200 205
Thr Cys Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr
210 215 220
Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu
225 230 235 240
Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys
245 250 255
Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser
260 265 270
Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln
275 280 285
Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp
290 295 300
Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His
305 310 315 320
Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro
325 330 335
Gly Lys
<210> 215
<211> 358
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine with signal sequence
IgG-B Fc protein A + C1q + CD16+ L (23) F (F00)
<400> 215
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Phe Pro Ala Ser Val Gln Leu His Glu Ala
20 25 30
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe
100 105 110
Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
115 120 125
Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu
130 135 140
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu
145 150 155 160
Phe Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp
165 170 175
Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln
180 185 190
Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr
195 200 205
Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys
210 215 220
Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala Leu Pro Ser Pro
225 230 235 240
Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser
245 250 255
Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val
260 265 270
Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val
275 280 285
Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr
290 295 300
Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
305 310 315 320
Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys
325 330 335
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu
340 345 350
Ser His Ser Pro Gly Lys
355
<210> 216
<211> 336
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine IgG-B Fc
Protein A + C1q + CD16+ L (23) F (F00)
<400> 216
Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
100 105 110
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
115 120 125
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Phe Ile Ala Arg Thr Pro
130 135 140
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
145 150 155 160
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
165 170 175
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
180 185 190
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
195 200 205
Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
210 215 220
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
225 230 235 240
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
245 250 255
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
260 265 270
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
275 280 285
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
290 295 300
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
305 310 315 320
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
325 330 335
<210> 217
<211> 384
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine with signal sequence
IgG-B Fc protein A + C1q + CD16+ L (23) Y (Y00)
<400> 217
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro
115 120 125
Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr
130 135 140
Ser Gly Asp Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
145 150 155 160
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
165 170 175
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile Ala Arg Thr Pro
180 185 190
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
195 200 205
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
210 215 220
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
225 230 235 240
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
245 250 255
Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
260 265 270
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
275 280 285
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
290 295 300
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
305 310 315 320
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
325 330 335
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
340 345 350
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
355 360 365
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
370 375 380
<210> 218
<211> 362
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine IgG-B Fc
Protein A + C1q + CD16+ L (23) Y (Y00)
<400> 218
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp Ser Gly Gly
115 120 125
Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala
130 135 140
Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro
145 150 155 160
Lys Asp Thr Leu Tyr Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val
165 170 175
Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val
180 185 190
Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln
195 200 205
Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln
210 215 220
Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala
225 230 235 240
Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala
245 250 255
His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser
260 265 270
Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro
275 280 285
Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser
290 295 300
Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe
305 310 315 320
Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp
325 330 335
Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr
340 345 350
Gln Glu Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 219
<211> 360
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine with signal sequence
IgG-B Fc protein A + C1q + CD16+ L (23) Y (Y00)
<400> 219
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu
130 135 140
Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
145 150 155 160
Thr Leu Tyr Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
165 170 175
Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly
180 185 190
Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn
195 200 205
Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp
210 215 220
Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala Leu Pro
225 230 235 240
Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln
245 250 255
Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn
260 265 270
Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile
275 280 285
Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr
290 295 300
Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr
305 310 315 320
Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe
325 330 335
Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu
340 345 350
Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 220
<211> 338
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-B Fc
Protein A + C1q + CD16+ L (23) Y (Y00)
<400> 220
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro
100 105 110
Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser
115 120 125
Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile Ala Arg
130 135 140
Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro
145 150 155 160
Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala
165 170 175
Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val
180 185 190
Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe
195 200 205
Thr Cys Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr
210 215 220
Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu
225 230 235 240
Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys
245 250 255
Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser
260 265 270
Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln
275 280 285
Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp
290 295 300
Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His
305 310 315 320
Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro
325 330 335
Gly Lys
<210> 221
<211> 358
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine with signal sequence
IgG-B Fc protein A + C1q + CD16+ L (23) Y (Y00)
<400> 221
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Phe Pro Ala Ser Val Gln Leu His Glu Ala
20 25 30
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe
100 105 110
Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
115 120 125
Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu
130 135 140
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu
145 150 155 160
Tyr Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp
165 170 175
Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln
180 185 190
Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr
195 200 205
Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys
210 215 220
Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala Leu Pro Ser Pro
225 230 235 240
Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser
245 250 255
Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val
260 265 270
Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val
275 280 285
Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr
290 295 300
Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
305 310 315 320
Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys
325 330 335
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu
340 345 350
Ser His Ser Pro Gly Lys
355
<210> 222
<211> 336
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine IgG-B Fc
Protein A + C1q + CD16+ L (23) Y (Y00)
<400> 222
Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
100 105 110
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
115 120 125
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile Ala Arg Thr Pro
130 135 140
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
145 150 155 160
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
165 170 175
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
180 185 190
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
195 200 205
Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
210 215 220
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
225 230 235 240
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
245 250 255
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
260 265 270
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
275 280 285
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
290 295 300
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
305 310 315 320
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
325 330 335
<210> 223
<211> 384
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine with signal sequence
IgG-B Fc protein A + C1Q + CD16+ Q (82) Y (0Y0)
<400> 223
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro
115 120 125
Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr
130 135 140
Ser Gly Asp Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
145 150 155 160
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
165 170 175
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
180 185 190
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
195 200 205
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
210 215 220
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
225 230 235 240
Leu Pro Ile Gly His Tyr Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
245 250 255
Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
260 265 270
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
275 280 285
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
290 295 300
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
305 310 315 320
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
325 330 335
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
340 345 350
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
355 360 365
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
370 375 380
<210> 224
<211> 362
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine IgG-B Fc
Protein A + C1Q + CD16+ Q (82) Y (0Y0)
<400> 224
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp Ser Gly Gly
115 120 125
Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala
130 135 140
Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro
145 150 155 160
Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val
165 170 175
Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val
180 185 190
Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln
195 200 205
Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Tyr
210 215 220
Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala
225 230 235 240
Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala
245 250 255
His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser
260 265 270
Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro
275 280 285
Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser
290 295 300
Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe
305 310 315 320
Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp
325 330 335
Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr
340 345 350
Gln Glu Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 225
<211> 360
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine with signal sequence
IgG-B Fc protein A + C1Q + CD16+ Q (82) Y (0Y0)
<400> 225
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu
130 135 140
Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
145 150 155 160
Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
165 170 175
Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly
180 185 190
Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn
195 200 205
Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Tyr Asp Trp
210 215 220
Leu Lys Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala Leu Pro
225 230 235 240
Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln
245 250 255
Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn
260 265 270
Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile
275 280 285
Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr
290 295 300
Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr
305 310 315 320
Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe
325 330 335
Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu
340 345 350
Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 226
<211> 338
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-B Fc
Protein A + C1Q + CD16+ Q (82) Y (0Y0)
<400> 226
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro
100 105 110
Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser
115 120 125
Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg
130 135 140
Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro
145 150 155 160
Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala
165 170 175
Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val
180 185 190
Ser Val Leu Pro Ile Gly His Tyr Asp Trp Leu Lys Gly Lys Gln Phe
195 200 205
Thr Cys Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr
210 215 220
Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu
225 230 235 240
Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys
245 250 255
Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser
260 265 270
Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln
275 280 285
Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp
290 295 300
Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His
305 310 315 320
Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro
325 330 335
Gly Lys
<210> 227
<211> 358
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine with signal sequence
IgG-B Fc protein A + C1Q + CD16+ Q (82) Y (0Y0)
<400> 227
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Phe Pro Ala Ser Val Gln Leu His Glu Ala
20 25 30
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe
100 105 110
Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
115 120 125
Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu
130 135 140
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu
145 150 155 160
Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp
165 170 175
Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln
180 185 190
Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr
195 200 205
Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Tyr Asp Trp Leu Lys
210 215 220
Gly Lys Gln Phe Thr Cys Lys Val Asn Asn Lys Ala Leu Pro Ser Pro
225 230 235 240
Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser
245 250 255
Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val
260 265 270
Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val
275 280 285
Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr
290 295 300
Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
305 310 315 320
Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys
325 330 335
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu
340 345 350
Ser His Ser Pro Gly Lys
355
<210> 228
<211> 336
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine IgG-B Fc
Protein A + C1Q + CD16+ Q (82) Y (0Y0)
<400> 228
Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
100 105 110
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
115 120 125
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
130 135 140
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
145 150 155 160
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
165 170 175
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
180 185 190
Leu Pro Ile Gly His Tyr Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
195 200 205
Lys Val Asn Asn Lys Ala Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
210 215 220
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
225 230 235 240
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
245 250 255
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
260 265 270
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
275 280 285
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
290 295 300
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
305 310 315 320
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
325 330 335
<210> 229
<211> 384
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-K (93) R K (97) I
A(98)G L(23)F (F00)
<400> 229
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro
115 120 125
Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr
130 135 140
Ser Gly Asp Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
145 150 155 160
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
165 170 175
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Phe Ile Ala Arg Thr Pro
180 185 190
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
195 200 205
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
210 215 220
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
225 230 235 240
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
245 250 255
Arg Val Asn Asn Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
260 265 270
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
275 280 285
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
290 295 300
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
305 310 315 320
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
325 330 335
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
340 345 350
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
355 360 365
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
370 375 380
<210> 230
<211> 362
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine IgG-B Fc
Protein A + C1q-CD16-K (93) R K (97) I A (98) G L (23) F (F00)
<400> 230
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp Ser Gly Gly
115 120 125
Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala
130 135 140
Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro
145 150 155 160
Lys Asp Thr Leu Phe Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val
165 170 175
Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val
180 185 190
Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln
195 200 205
Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln
210 215 220
Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Ile Gly
225 230 235 240
Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala
245 250 255
His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser
260 265 270
Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro
275 280 285
Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser
290 295 300
Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe
305 310 315 320
Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp
325 330 335
Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr
340 345 350
Gln Glu Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 231
<211> 360
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-K (93) R K (97) I
A(98)G L(23)F (F00)
<400> 231
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu
130 135 140
Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
145 150 155 160
Thr Leu Phe Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
165 170 175
Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly
180 185 190
Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn
195 200 205
Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp
210 215 220
Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Ile Gly Leu Pro
225 230 235 240
Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln
245 250 255
Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn
260 265 270
Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile
275 280 285
Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr
290 295 300
Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr
305 310 315 320
Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe
325 330 335
Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu
340 345 350
Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 232
<211> 338
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-B Fc
Protein A + C1q-CD16-K (93) R K (97) I A (98) G L (23) F (F00)
<400> 232
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro
100 105 110
Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser
115 120 125
Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Phe Ile Ala Arg
130 135 140
Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro
145 150 155 160
Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala
165 170 175
Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val
180 185 190
Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe
195 200 205
Thr Cys Arg Val Asn Asn Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr
210 215 220
Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu
225 230 235 240
Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys
245 250 255
Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser
260 265 270
Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln
275 280 285
Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp
290 295 300
Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His
305 310 315 320
Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro
325 330 335
Gly Lys
<210> 233
<211> 358
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-K (93) R K (97) I
A(98)G L(23)F (F00)
<400> 233
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Phe Pro Ala Ser Val Gln Leu His Glu Ala
20 25 30
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe
100 105 110
Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
115 120 125
Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu
130 135 140
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu
145 150 155 160
Phe Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp
165 170 175
Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln
180 185 190
Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr
195 200 205
Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys
210 215 220
Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Ile Gly Leu Pro Ser Pro
225 230 235 240
Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser
245 250 255
Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val
260 265 270
Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val
275 280 285
Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr
290 295 300
Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
305 310 315 320
Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys
325 330 335
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu
340 345 350
Ser His Ser Pro Gly Lys
355
<210> 234
<211> 336
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine IgG-B Fc
Protein A + C1q-CD16-K (93) R K (97) I A (98) G L (23) F (F00)
<400> 234
Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
100 105 110
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
115 120 125
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Phe Ile Ala Arg Thr Pro
130 135 140
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
145 150 155 160
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
165 170 175
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
180 185 190
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
195 200 205
Arg Val Asn Asn Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
210 215 220
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
225 230 235 240
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
245 250 255
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
260 265 270
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
275 280 285
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
290 295 300
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
305 310 315 320
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
325 330 335
<210> 235
<211> 384
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-K (93) R K (97) I
A(98)G L(23)Y (Y00)
<400> 235
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro
115 120 125
Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr
130 135 140
Ser Gly Asp Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
145 150 155 160
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
165 170 175
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile Ala Arg Thr Pro
180 185 190
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
195 200 205
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
210 215 220
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
225 230 235 240
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
245 250 255
Arg Val Asn Asn Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
260 265 270
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
275 280 285
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
290 295 300
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
305 310 315 320
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
325 330 335
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
340 345 350
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
355 360 365
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
370 375 380
<210> 236
<211> 362
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine IgG-B Fc
Protein A + C1q-CD16-K (93) R K (97) I A (98) G L (23) Y (Y00)
<400> 236
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp Ser Gly Gly
115 120 125
Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala
130 135 140
Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro
145 150 155 160
Lys Asp Thr Leu Tyr Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val
165 170 175
Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val
180 185 190
Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln
195 200 205
Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln
210 215 220
Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Ile Gly
225 230 235 240
Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala
245 250 255
His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser
260 265 270
Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro
275 280 285
Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser
290 295 300
Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe
305 310 315 320
Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp
325 330 335
Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr
340 345 350
Gln Glu Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 237
<211> 360
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-K (93) R K (97) I
A(98)G L(23)Y (Y00)
<400> 237
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu
130 135 140
Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
145 150 155 160
Thr Leu Tyr Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
165 170 175
Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly
180 185 190
Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn
195 200 205
Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp
210 215 220
Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Ile Gly Leu Pro
225 230 235 240
Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln
245 250 255
Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn
260 265 270
Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile
275 280 285
Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr
290 295 300
Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr
305 310 315 320
Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe
325 330 335
Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu
340 345 350
Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 238
<211> 338
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-B Fc
Protein A + C1q-CD16-K (93) R K (97) I A (98) G L (23) Y (Y00)
<400> 238
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro
100 105 110
Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser
115 120 125
Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile Ala Arg
130 135 140
Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro
145 150 155 160
Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala
165 170 175
Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val
180 185 190
Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe
195 200 205
Thr Cys Arg Val Asn Asn Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr
210 215 220
Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu
225 230 235 240
Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys
245 250 255
Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser
260 265 270
Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln
275 280 285
Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp
290 295 300
Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His
305 310 315 320
Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro
325 330 335
Gly Lys
<210> 239
<211> 358
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-K (93) R K (97) I
A(98)G L(23)Y (Y00)
<400> 239
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Phe Pro Ala Ser Val Gln Leu His Glu Ala
20 25 30
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe
100 105 110
Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
115 120 125
Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu
130 135 140
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu
145 150 155 160
Tyr Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp
165 170 175
Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln
180 185 190
Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr
195 200 205
Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Gln Asp Trp Leu Lys
210 215 220
Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Ile Gly Leu Pro Ser Pro
225 230 235 240
Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser
245 250 255
Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val
260 265 270
Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val
275 280 285
Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr
290 295 300
Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
305 310 315 320
Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys
325 330 335
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu
340 345 350
Ser His Ser Pro Gly Lys
355
<210> 240
<211> 336
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine IgG-B Fc
Protein A + C1q-CD16-K (93) R K (97) I A (98) G L (23) F (Y00)
<400> 240
Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
100 105 110
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
115 120 125
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile Ala Arg Thr Pro
130 135 140
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
145 150 155 160
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
165 170 175
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
180 185 190
Leu Pro Ile Gly His Gln Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
195 200 205
Arg Val Asn Asn Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
210 215 220
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
225 230 235 240
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
245 250 255
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
260 265 270
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
275 280 285
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
290 295 300
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
305 310 315 320
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
325 330 335
<210> 241
<211> 384
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-K (93) R K (97) I
A(98)G Q(82)Y (0Y0)
<400> 241
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Phe Glu Phe Asn Pro
115 120 125
Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp Thr Asn Ser Thr
130 135 140
Ser Gly Asp Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
145 150 155 160
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
165 170 175
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
180 185 190
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
195 200 205
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
210 215 220
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
225 230 235 240
Leu Pro Ile Gly His Tyr Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
245 250 255
Arg Val Asn Asn Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
260 265 270
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
275 280 285
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
290 295 300
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
305 310 315 320
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
325 330 335
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
340 345 350
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
355 360 365
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
370 375 380
<210> 242
<211> 362
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 2-variant canine IgG-B Fc
Protein A + C1q-CD16-K (93) R K (97) I A (98) G Q (82) Y (0Y0)
<400> 242
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Phe Glu Phe Asn Pro Glu Asp Pro Ile Pro Val
100 105 110
Ser Phe Ser Pro Val Asp Thr Asn Ser Thr Ser Gly Asp Ser Gly Gly
115 120 125
Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala
130 135 140
Pro Glu Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro
145 150 155 160
Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val
165 170 175
Val Asp Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val
180 185 190
Asp Gly Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln
195 200 205
Phe Asn Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Tyr
210 215 220
Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Ile Gly
225 230 235 240
Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala
245 250 255
His Gln Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser
260 265 270
Lys Asn Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro
275 280 285
Asp Ile Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser
290 295 300
Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe
305 310 315 320
Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp
325 330 335
Thr Phe Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr
340 345 350
Gln Glu Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 243
<211> 360
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-B Fc with signal sequence
Protein A + C1q-CD16-K (93) R K (97) I
A(98)G Q(82)Y (0Y0)
<400> 243
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Val Ser Phe Pro Ala Ser Val Gln Leu His
20 25 30
Glu Ala Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly
35 40 45
Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn
50 55 60
Glu Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu
65 70 75 80
Thr Val Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn
85 90 95
Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala
100 105 110
Ala Phe Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser
115 120 125
Gly Gly Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu
130 135 140
Met Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
145 150 155 160
Thr Leu Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
165 170 175
Leu Asp Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly
180 185 190
Lys Gln Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn
195 200 205
Gly Thr Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Tyr Asp Trp
210 215 220
Leu Lys Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Ile Gly Leu Pro
225 230 235 240
Ser Pro Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln
245 250 255
Pro Ser Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn
260 265 270
Thr Val Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile
275 280 285
Asp Val Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr
290 295 300
Arg Thr Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr
305 310 315 320
Ser Lys Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe
325 330 335
Ile Cys Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu
340 345 350
Ser Leu Ser His Ser Pro Gly Lys
355 360
<210> 244
<211> 338
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 3-variant canine IgG-B Fc
Protein A + C1q-CD16-K (93) R K (97) I A (98) G Q (82) Y (0Y0)
<400> 244
Val Ser Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His
1 5 10 15
His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu
20 25 30
Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe
35 40 45
Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys
50 55 60
Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu
65 70 75 80
Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala
85 90 95
Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro
100 105 110
Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser
115 120 125
Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg
130 135 140
Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro
145 150 155 160
Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala
165 170 175
Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val
180 185 190
Ser Val Leu Pro Ile Gly His Tyr Asp Trp Leu Lys Gly Lys Gln Phe
195 200 205
Thr Cys Arg Val Asn Asn Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr
210 215 220
Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu
225 230 235 240
Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys
245 250 255
Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser
260 265 270
Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln
275 280 285
Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp
290 295 300
Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His
305 310 315 320
Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro
325 330 335
Gly Lys
<210> 245
<211> 358
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine with signal sequence
IgG-B Fc protein A + C1q-CD16-K (93) R K (97) I
A(98)G Q(82)Y (0Y0)
<400> 245
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Leu Arg Gly Ala Arg Cys Phe Pro Ala Ser Val Gln Leu His Glu Ala
20 25 30
Val Glu Leu His His Trp Cys Ile Pro Phe Ser Val Asp Gly Gln Pro
35 40 45
Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser Val Leu Asn Glu Thr
50 55 60
Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Val Ala Asn Glu Thr Val
65 70 75 80
Arg His Gly Cys Leu Arg Leu Asn Gln Pro Thr His Val Asn Asn Gly
85 90 95
Asn Tyr Thr Leu Leu Ala Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe
100 105 110
Val Met Ala Ala Phe Met Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly
115 120 125
Gly Ser Arg Pro Pro Asp Cys Pro Lys Cys Pro Ala Pro Glu Met Leu
130 135 140
Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu
145 150 155 160
Leu Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Leu Asp
165 170 175
Pro Glu Asp Pro Glu Val Gln Ile Ser Trp Phe Val Asp Gly Lys Gln
180 185 190
Met Gln Thr Ala Lys Thr Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr
195 200 205
Tyr Arg Val Val Ser Val Leu Pro Ile Gly His Tyr Asp Trp Leu Lys
210 215 220
Gly Lys Gln Phe Thr Cys Arg Val Asn Asn Ile Gly Leu Pro Ser Pro
225 230 235 240
Ile Glu Arg Thr Ile Ser Lys Ala Arg Gly Gln Ala His Gln Pro Ser
245 250 255
Val Tyr Val Leu Pro Pro Ser Arg Glu Glu Leu Ser Lys Asn Thr Val
260 265 270
Ser Leu Thr Cys Leu Ile Lys Asp Phe Phe Pro Pro Asp Ile Asp Val
275 280 285
Glu Trp Gln Ser Asn Gly Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr
290 295 300
Thr Pro Pro Gln Leu Asp Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys
305 310 315 320
Leu Ser Val Asp Lys Ser Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys
325 330 335
Ala Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Glu Ser Leu
340 345 350
Ser His Ser Pro Gly Lys
355
<210> 246
<211> 336
<212> PRT
<213> Artificial sequence
<220>
<223> synthetic exemplary canine TrkA ECD v 4-variant canine IgG-B Fc
Protein A + C1q-CD16-K (93) R K (97) I A (98) G Q (82) Y (0Y0)
<400> 246
Phe Pro Ala Ser Val Gln Leu His Glu Ala Val Glu Leu His His Trp
1 5 10 15
Cys Ile Pro Phe Ser Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp
20 25 30
Leu Phe Asn Gly Ser Val Leu Asn Glu Thr Ser Phe Ile Phe Thr Glu
35 40 45
Phe Leu Glu Pro Val Ala Asn Glu Thr Val Arg His Gly Cys Leu Arg
50 55 60
Leu Asn Gln Pro Thr His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala
65 70 75 80
Ala Asn Pro Ser Gly Arg Ala Ala Ala Phe Val Met Ala Ala Phe Met
85 90 95
Asp Asn Pro Ser Gly Gly Gly Ser Gly Gly Gly Ser Arg Pro Pro Asp
100 105 110
Cys Pro Lys Cys Pro Ala Pro Glu Met Leu Gly Gly Pro Ser Val Phe
115 120 125
Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu Leu Ile Ala Arg Thr Pro
130 135 140
Glu Val Thr Cys Val Val Val Asp Leu Asp Pro Glu Asp Pro Glu Val
145 150 155 160
Gln Ile Ser Trp Phe Val Asp Gly Lys Gln Met Gln Thr Ala Lys Thr
165 170 175
Gln Pro Arg Glu Glu Gln Phe Asn Gly Thr Tyr Arg Val Val Ser Val
180 185 190
Leu Pro Ile Gly His Tyr Asp Trp Leu Lys Gly Lys Gln Phe Thr Cys
195 200 205
Arg Val Asn Asn Ile Gly Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser
210 215 220
Lys Ala Arg Gly Gln Ala His Gln Pro Ser Val Tyr Val Leu Pro Pro
225 230 235 240
Ser Arg Glu Glu Leu Ser Lys Asn Thr Val Ser Leu Thr Cys Leu Ile
245 250 255
Lys Asp Phe Phe Pro Pro Asp Ile Asp Val Glu Trp Gln Ser Asn Gly
260 265 270
Gln Gln Glu Pro Glu Ser Lys Tyr Arg Thr Thr Pro Pro Gln Leu Asp
275 280 285
Glu Asp Gly Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Asp Lys Ser
290 295 300
Arg Trp Gln Arg Gly Asp Thr Phe Ile Cys Ala Val Met His Glu Ala
305 310 315 320
Leu His Asn His Tyr Thr Gln Glu Ser Leu Ser His Ser Pro Gly Lys
325 330 335
- 上一篇：一种医用注射器针头装配设备
- 下一篇：可活化白介素-2多肽及其使用方法