阅读说明：本技术 基于精子相关蛋白catsper的避孕疫苗 (Contraceptive vaccine based on sperm-associated protein CATSPER ) 是由 C·比斯托夫 J-J·L·成 于 2020-02-10 设计创作，主要内容包括：一种包含避孕嵌合病毒样颗粒的组合物,其具有抗原运载体结构域和该抗原运载体结构域中的来自精细胞的一种或多种抗原区域,其中所述抗原运载体结构域包括人乳头瘤病毒L1衣壳蛋白,所述抗原区域包括Catsper离子通道复合物的一种或多种结构元件。当给予接受者时,避孕疫苗刺激抗精子抗体的产生,所述抗体结合精细胞后会抑制精细胞的运动,从而抑制精细胞使卵细胞受精的能力。通过给接受者过量使用逆转剂可以在短时间或延长时间内逆转组合物诱导的免疫不孕,所述逆转剂缺乏抗原运载体结构域,但具有与所述一种或多种抗原区域基本相同的蛋白质序列以隔离抗精子抗体。(A composition comprising a contraceptive chimeric virus-like particle having an antigen carrier domain and one or more antigenic regions from sperm cells in the antigen carrier domain, wherein the antigen carrier domain comprises human papilloma virus L1 capsid protein, said antigenic regions comprising one or more structural elements of a cats ion channel complex. When administered to a recipient, the contraceptive vaccine stimulates the production of anti-sperm antibodies that, upon binding to sperm cells, inhibit the motility of the sperm cells, thereby inhibiting the ability of the sperm cells to fertilize an egg cell. The immune infertility induced by the composition can be reversed over a short or extended period of time by overdosing the recipient with a reversing agent that lacks the antigen-carrier domain but has substantially the same protein sequence as the one or more antigenic domains to sequester the anti-sperm antibodies.)

1. A contraceptive chimeric virus-like particle comprising:

an antigen carrier domain; and

one or more antigenic regions from sperm cells within the antigen carrier domain.

2. The chimeric virus-like particle of claim 1, wherein the antigen carrier domain comprises one or more capsid proteins.

3. The chimeric virus-like particle of claim 2, wherein the one or more capsid proteins comprise L1 from human papilloma virus.

4. The chimeric virus-like particle of claim 1, wherein CGP amino acid residues are adjacent to the N-terminus and GPC amino acid residues are adjacent to the C-terminus of the one or more antigenic regions.

5. The chimeric virus-like particle of claim 1, wherein the one or more antigenic regions comprise structural elements of a Catsper ion channel complex.

6. The chimeric virus-like particle of claim 5, wherein the structural element comprises at least a portion of one or more loops located between transmembrane helix segments of the Catsper ion channel complex.

7. The chimeric virus-like particle of claim 6, wherein the structural element comprises at least a portion of: the loop between Catsper1s1 and Catsper1s 2, the loop between Catsper2 s5 and Catsper2 p-loop, the loop between Catsper1s 3 and Catsper1s 4, the loop between Catsper2 s1 and Catsper2 s2, the loop between Catsper3 s3 and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 p-loop, the loop between Catsper3 p-loop and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 p-loop, the loop between Catsper3 s3 and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 s3, the combination thereof.

8. The chimeric virus-like particle of claim 7, wherein the virus-like particle comprises 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, or a combination thereof.

9. A method of making a contraceptive chimeric virus-like particle comprising:

inserting a gene for an antigen carrier protein into a plasmid;

preparing overlapping primers for the antigen carrier and the chimeric gene from one or more antigenic regions of sperm cells;

performing a polymerase chain reaction to amplify the chimeric gene; and

synthesizing virus-like particles from the chimeric gene.

10. The method of claim 9, wherein the one or more antigenic regions comprise a structural element of the Catsper ion channel complex.

11. The method of claim 10, wherein the structural element comprises at least a portion of one or more loops located between transmembrane helix segments of the Catsper ion channel complex.

12. The method of claim 11, wherein the structural element comprises at least part of: the loop between Catsper1s1 and Catsper1s 2, the loop between Catsper2 s5 and Catsper2 p-loop, the loop between Catsper1s 3 and Catsper1s 4, the loop between Catsper2 s1 and Catsper2 s2, the loop between Catsper3 s3 and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 p-loop, the loop between Catsper3 p-loop and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 p-loop, the loop between Catsper3 s3 and Catsper3 s3, the combination thereof.

13. The method of claim 12, wherein the virus-like particle comprises 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, or a combination thereof.

14. A method for providing a contraceptive treatment to a recipient, comprising:

preparing a composition comprising a contraceptive chimeric virus-like particle comprising an antigen-carrier domain and one or more antigenic regions from sperm cells within the antigen-carrier domain; and

administering the composition to a recipient to enhance the recipient's immune response to the one or more antigenic regions.

15. The method of claim 14, wherein the antigen carrier domain comprises L1 from human papilloma virus.

16. The method of claim 14, wherein the one or more antigenic regions comprise a structural element of the Catsper ion channel complex, wherein the structural element comprises at least a portion of: the loop between Catsper1s1 and Catsper1s 2, the loop between Catsper2 s5 and Catsper2 p-loop, the loop between Catsper1s 3 and Catsper1s 4, the loop between Catsper2 s1 and Catsper2 s2, the loop between Catsper3 s3 and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 p-loop, the loop between Catsper3 p-loop and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 p-loop, the loop between Catsper3 s3 and Catsper3 s3, the combination thereof.

17. The method of claim 16, wherein the one or more antigenic regions comprise seq.d.no. 8, seq.d.no. 9, seq.d.no. 10, seq.d.no. 11, seq.d.no. 12, seq.d.no. 13, seq.d.no. 14, seq.d.no. 15, seq.d.no. 16, seq.d.no. 17, seq.d.no. 18, seq.d.no. 19, seq.d.no. 20, seq.d.no. 21, seq.d.no. 22, or combinations thereof.

18. The method of claim 14, wherein the composition is administered subcutaneously, intravenously, intranasally, or a combination thereof.

19. The method of claim 14, further comprising:

administering to the recipient a supplemental composition to reverse the effect of the composition, the supplemental composition comprising a reversing agent having a protein sequence substantially identical to the one or more antigenic regions.

20. The method of claim 19, wherein the reversing agent comprises one or more peptides comprising 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. 16, seq id No. 17, seq id No. 18, seq id No. 19, seq id No. 20, seq id No. 21, seq id No. 22, or combinations thereof.

Background

Almost half of the world's pregnancy is unexpected or untimely. Current long-lasting reversible contraceptive techniques focus on hormonal approaches or surgery. Despite their overall effectiveness, contraceptive techniques available to both females and males suffer from a myriad of disadvantages. As an example, some women are unable to tolerate hormonal contraception. In addition, hormonal approaches often require strict discipline or discomfort of the recipient. For example, pill or patch regimens may be required to be taken or administered periodically during the time that contraceptive efficacy is desired. Implantable or intrauterine devices may cause mild to severe side effects and complications, and may still require clinical visits for reversal or re-implantation.

Thus, there is a need for an effective, durable and reversible contraceptive treatment that is more convenient to use while also reducing the burden associated with treatment for the recipient in need of contraceptive effect.

Disclosure of Invention

Accordingly, some embodiments of the present disclosure relate to a contraceptive chimeric virus-like particle comprising an antigen carrier domain and one or more antigenic regions from sperm cells within the antigen carrier domain. In some embodiments, the antigen carrier domain comprises one or more capsid proteins. In some embodiments, the one or more capsid proteins comprise L1 from human papilloma virus. In some embodiments, the CGP amino acid residues are adjacent to the N-terminus of the one or more antigenic regions and the GPC amino acid residues are adjacent to the C-terminus of the one or more antigenic regions. In some embodiments, the one or more antigenic regions comprise a structural element of a Catsper ion channel complex. In some embodiments, the structural element comprises at least a portion of one or more loops located between transmembrane helix segments of a Catsper ion channel complex. In some embodiments, the structural element comprises at least part of: the loop between Catsper1s1 and Catsper1s 2, the loop between Catsper2 s5 and Catsper2 p-loop, the loop between Catsper1s 3 and Catsper1s 4, the loop between Catsper2 s1 and Catsper2 s2, the loop between Catsper3 s3 and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 p-loop, the loop between Catsper3 p-loop and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 p-loop, the loop between Catsper3 s3 and Catsper3 s3, the combination thereof. In some embodiments, the virus-like particle comprises 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, or a combination thereof.

Some embodiments of the present disclosure relate to a method of making a contraceptive chimeric virus-like particle comprising inserting a gene of an antigen carrier protein into a plasmid, making overlapping primers for the antigen carrier and the chimeric gene from one or more antigenic regions of sperm cells, performing a polymerase chain reaction to amplify the chimeric gene, and synthesizing the virus-like particle from the chimeric gene. In some embodiments, the one or more antigenic regions comprise a structural element of a Catsper ion channel complex. In some embodiments, the structural element comprises at least a portion of one or more loops located between transmembrane helix segments of a Catsper ion channel complex. In some embodiments, the structural element comprises at least part of: the loop between Catsper1s1 and Catsper1s 2, the loop between Catsper2 s5 and Catsper2 p-loops, the loop between Catsper1s 3 and Catsper1s 4, the loop between Catsper2 s1 and Catsper2 s2, the loop between Catsper3 s1 and Catsper3 s2, the loop between Catsper2 s2 and Catsper2 p-loops, the loop between Catsper2 p-loops and Catsper2 s2, the loop between Catsper2 s2 and Catsper2 p-loops, the loop between Catsper2 and Catsper 4672 s2, the loop between Catsper2 s2 and Catsper2 s2, the combination thereof. In some embodiments, the virus-like particle comprises 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, or a combination thereof.

Some embodiments of the present disclosure relate to a method of providing a contraceptive treatment to a recipient, comprising preparing a composition comprising a contraceptive chimeric virus-like particle comprising an antigen carrier domain and one or more antigenic regions from a sperm cell in the antigen carrier domain, and administering the composition to the recipient to enhance the recipient's immune response to the one or more antigenic regions. In some embodiments, the antigen carrier domain comprises L1 from human papilloma virus. In some embodiments, the one or more antigenic regions comprise a structural element of a Catsper ion channel complex, wherein the structural element comprises at least a portion of: the loop between Catsper1s1 and Catsper1s 2, the loop between Catsper2 s5 and Catsper2 p-loop, the loop between Catsper1s 3 and Catsper1s 4, the loop between Catsper2 s1 and Catsper2 s2, the loop between Catsper3 s3 and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 p-loop, the loop between Catsper3 p-loop and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 p-loop, the loop between Catsper3 s3 and Catsper3 s3, the combination thereof. In some embodiments, the one or more antigenic regions comprise 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. 16, seq id No. 17, seq id No. 18, seq id No. 19, seq id No. 20, seq id No. 21, seq id No. 22, or combinations thereof. In some embodiments, the composition is administered subcutaneously, intravenously, intranasally, or a combination thereof. In some embodiments, the method further comprises administering to the recipient a supplemental composition comprising a reversing agent having a protein sequence substantially identical to the one or more antigenic regions to reverse the effects of the aforementioned composition. In some embodiments, the reversal agent comprises one or more peptides comprising 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. 16, seq id No. 17, seq id No. 18, seq id No. 19, seq id No. 20, seq id No. 21, seq id No. 22, or combinations thereof.

Brief description of the drawings

For the purpose of illustrating the invention, the drawings show embodiments of the disclosed subject matter. It should be understood, however, that the application is not limited to the precise arrangements and instrumentalities shown in the drawings, wherein:

fig. 1 is a schematic illustration of a contraceptive chimeric virus-like particle according to some embodiments of the present disclosure;

FIG. 2 is a diagram of a method of making a contraceptive chimeric virus-like particle according to some embodiments of the present disclosure;

fig. 3A is a diagram of a method of providing a contraceptive treatment to a recipient according to some embodiments of the present disclosure; and

fig. 3B is a diagram of a method of providing a contraceptive treatment to a recipient according to some embodiments of the present disclosure.

Detailed Description

Referring now to fig. 1, some embodiments of the present disclosure relate to a vaccine for providing a contraceptive treatment to an individual, user, recipient, or the like. In some embodiments, the contraceptive vaccine provides contraceptive benefits to the user that are substantially equivalent in effectiveness to traditional hormonal contraception, implant contraception, physical contraception, and the like. In some embodiments, the contraceptive vaccine enhances the immune response of the user to structures present on the sperm cells. In some embodiments, the contraceptive vaccine enhances the immune response of the user to the structure of the protein or combination of proteins present on the sperm cells. In some embodiments, the contraceptive vaccine stimulates the production of anti-sperm antibodies in the user. In some embodiments, the contraceptive vaccine stimulates the production of anti-sperm antibodies that bind to sperm cells that inhibit the sperm cells' ability to fertilize an egg cell. In some embodiments, the contraceptive vaccine stimulates the production of anti-sperm antibodies that, upon binding to sperm cells, inhibit the motility of the sperm cells. Without wishing to be bound by theory, when administered to women, contraceptive vaccines stimulate the production of anti-sperm antibodies, e.g., in the vaginal mucosa, which bind to and prevent the hyperactive movement of sperm, rendering sperm unable to penetrate the ovum. Also without wishing to be bound by theory, when administered to men, contraceptive vaccines stimulate the production of anti-sperm antibodies, e.g., in the epididymis, which render the sperm substantially inert.

In some embodiments, a contraceptive vaccine comprises a contraceptive chimeric virus-like particle (VLP) 100. In some embodiments, VLP100 comprises two or more domains 102. In some embodiments, the VLP100 comprises an antigen carrier domain 104. In some embodiments, the antigen carrier domain 104 comprises one or more capsid proteins. In some embodiments, the one or more capsid proteins comprise L1 from human papilloma virus.

In some embodiments, VLP100 comprises antigen domain 106. In some embodiments, antigenic domain 106 includes one or more antigenic regions 106A. In some embodiments, the antigen region 106A is located within the antigen carrier domain 104. In some embodiments, the antigenic region 106A satisfies one or more of the following: exposed on or around the outer surface of the sperm cell, present only in the sperm cell and not in other tissues within the human body, and required for sperm function. In some embodiments, antigenic region 106A comprises a structural feature or a partial structural feature from a sperm cell. In some embodiments, antigenic region 106A comprises a structural element of a protein or combination of proteins present on a sperm cell. In some embodiments, the antigenic region 106A is present on a sperm cell flagellum. In some embodiments, antigenic region 106A comprises a structural element of a Catsper ion channel complex. In some embodiments, antigenic region 106A comprises at least a portion of one or more loops located at least between transmembrane helix segments of a Catsper ion channel complex. In some embodiments, the antigenic region 106A includes at least a portion of: the loop between Catsper1s1 and Catsper1s 2, the loop between Catsper2 s5 and Catsper2 p-loop, the loop between Catsper1s 3 and Catsper1s 4, the loop between Catsper2 s1 and Catsper2 s2, the loop between Catsper3 s3 and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 p-loop, the loop between Catsper3 p-loop and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 p-loop, the loop between Catsper3 s3 and Catsper3 s3, the combination thereof. In some embodiments, the antigen region 106A comprises 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. 16, seq id No. 17, seq id No. 18, seq id No. 19, seq id No. 20, seq id No. 21, seq id No. 22, or a combination thereof.

In some embodiments, the CGP amino acid residues are adjacent to the N-terminus of antigenic region 106A. In some embodiments, GPC amino acid residues are adjacent to the C-terminus of one or more antigenic regions 106A. Without intending to be limited by theory, these additional residues help stabilize the antigenic region 106A within the antigen carrier domain 104. In some embodiments, the VLP100 comprises 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, or combinations thereof.

In some embodiments, the VLP100 is comprised in a composition. In some embodiments, the composition includes one or more preservatives, stabilizers, wetting agents, emulsifiers, buffers, fillers, and the like. In some embodiments, the composition is formulated for administration to a user subcutaneously, intravenously, intranasally, or a combination thereof.

Referring now to fig. 2, some embodiments of the present disclosure relate to a method 200 of making a contraceptive chimeric VLP. At 202, the gene for the antigen carrier protein is inserted into a plasmid. At 204, overlapping primers for the antigen carrier and the chimeric gene from one or more antigenic regions of the sperm cell are prepared. As described above, in some embodiments, one or more antigenic regions comprise a structural element of a Catsper ion channel complex, e.g., a structural element comprising at least a portion of one or more loops located between transmembrane helix segments of the complex. In some embodiments, the structural element comprises at least part of: the loop between Catsper1s1 and Catsper1s 2, the loop between Catsper2 s5 and Catsper2 p-loop, the loop between Catsper1s 3 and Catsper1s 4, the loop between Catsper2 s1 and Catsper2 s2, the loop between Catsper3 s3 and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 p-loop, the loop between Catsper3 p-loop and Catsper3 s3, the loop between Catsper3 s3 and Catsper3 p-loop, the loop between Catsper3 s3 and Catsper3 s3, the combination thereof. At 206, a polymerase chain reaction is performed to amplify the chimeric gene. At 208, VLPs are synthesized from the chimeric gene. In some embodiments, the VLP is synthesized in bacterial or yeast cell culture. In some embodiments, the VLP comprises 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, or combinations thereof. In some embodiments, the VLP folds spontaneously. In some embodiments, the VLP is one or more of the VLPs 100 described above.

Referring now to fig. 3A, some embodiments of the present disclosure relate to a method 300 for providing a contraceptive treatment to a recipient. At 302, a composition is prepared comprising a contraceptive chimeric VLP comprising an antigen carrier domain and one or more antigenic regions from sperm cells within the antigen carrier domain. In some embodiments, the chimeric VLP is one or more of the VLPs 100 described above. At 304, the composition is administered to the recipient to enhance the recipient's immune response to the one or more antigenic regions. In some embodiments, the composition is administered to the recipient subcutaneously, intravenously, intranasally, or a combination thereof. Without intending to be limited by theory, contraceptive immunity is expected to last from about 1 year to about 9 years based on prior experience in immunizing women against HPV. Referring now to fig. 3B, in some embodiments, at 306, the recipient is administered a supplemental composition to reverse the effects of the aforementioned composition. In some embodiments, the supplemental composition comprises a reversal agent having a protein sequence substantially identical to the one or more antigenic regions. In some embodiments, the supplemental composition does not comprise an antigen carrier protein. In some embodiments, the reversal agent comprises one or more peptides comprising 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. 16, seq id No. 17, seq id No. 18, seq id No. 19, seq id No. 20, seq id No. 21, seq id No. 22, or combinations thereof. In some embodiments, the supplemental composition includes one or more preservatives, stabilizers, wetting agents, emulsifiers, buffers, fillers, and the like. Without wishing to be bound by theory, immune infertility induced by the composition is reversed by sequestering the antisperm antibodies using an excess of the supplemental composition. In some embodiments, if the sperm-immunized individual wishes to regain fertility for a short period of time, a fertility window may be established by application of a reversal agent.

Examples

Design of chimeric L1.For example, a chimeric L1 VLP-forming protein was prepared that included short antigen segments from the sperm cation channel, cats er. The antigenic segment of Catsper is generated byThe extracellular loops are identified as predicted using homology modeling. The site of insertion of the L1 gene was identified by multiple sequence alignment of homologous papillomavirus L1 sequences. Three candidate insertion sites were identified as sites for natural insertions/deletions. A sequence encompassing cgp. The DNA sequence was designed using DNAworks.

Cloning of chimeric L1.HPV type 11L 1 gene was synthesized using assembly PCR method. This gene was inserted into the NdeI and EcoRI cloning sites of the pET28a + vector. Plasmids were grown and purified from cell culture using the E.coli strain DH5 α. Chimeric constructs were prepared by amplifying plasmids using overlapping oligonucleotides containing the sequence of the desired antigenic region (inverse PCR). The amplicon was transformed into DH5 α cells for plasmid amplification and transferred into BL21(DE3) pLysS cells for protein expression.

Isolation of Inclusion Bodies (IB).The recombinant proteins HPV type 11L 1 protein (L1), L1 with Catsper loop S3-S4 (P1), and L1 with Catsper-epsilon loop 331-348 (C5) were expressed in E.coli. Recombinant proteins (L1, P1 and C5) were purified from Inclusion Bodies (IB) isolated from E.coli Bl21(DE3) transformed with the plasmid constructs pET28a-L1-P1 and pET28 a-L1-C5. After induction of recombinant protein synthesis, cells were harvested from 500ml of transformed E.coli cell culture. Approximately 1g weight of wet cell biomass was resuspended in 25ml of buffer (50mM Tris, 50mM NaCl, pH9.5) by gentle stirring. The suspension was sonicated (run for 10 seconds then stop for 10 seconds, 15 cycles in ice) and then centrifuged at 12,000g for 20 minutes at 4 ℃ to precipitate the IB. The harvested IB's were washed twice with wash buffer 50mM Tris (pH 9.5).

Dissolution and refolding.The relatively purified IB was dissolved in 5ml of solubilization buffer (50mM Tris (pH9.5), 50mM NaCl, 8M urea) and gently stirred at room temperature for 2 hours. The resulting suspension was centrifuged at 11000rpm for 36 minutes at 25 ℃ to collect total soluble protein. Solubilized IBs were refolded using a pulsed dilution method in 10 volumes of ice-cold refolding buffer (50mM Tris, 50mM NaCl, 5% beta-mercaptoethanol and 100mM L-arginine (pH9.5)) at a rate of about 0.1 ml/min. Refolded proteins at 11000 ℃Centrifuge at rpm for 45 minutes.

Dialysis and in vitro assembly of virus-like particles (VLPs).The refolded protein was filtered through a 0.2 μm filter (VWR) and dialyzed against dialysis buffer (50mM Tris, 100mM NaCl, pH9.5) at 4 ℃ using a 12-14kDa cut-off dialysis membrane (Spectra/Por, VWR). After 3 dialysis buffer changes at 3 hour intervals, the NaCl concentration was gradually increased to 500 mM. Final dialysis was performed for 2 hours against high salt buffer (50mM Tris, 500mM NaCl, pH 9.5).

Methods and systems according to some embodiments of the present disclosure relate to contraceptive vaccines for providing a sustained but reversible contraceptive effect. The structure of the contraceptive vaccine stimulates an immune response to antigens present only in the sperm cells, producing anti-sperm antibodies that effectively bind sperm and render these cells incapable of insemination. The contraceptive vaccine is simple to produce and use. The need for hormonal treatment and/or surgery is avoided because no action other than vaccination is required to achieve the benefits of the compositions of the present disclosure. Finally, contraceptive immunity is easily and immediately reversible, and over time immunity automatically reverts to the contraceptive state.

Although the present invention has been described and illustrated with reference to exemplary embodiments thereof, it should be understood by those skilled in the art that the foregoing and various other changes, omissions and additions may be made therein and thereto, without parting from the spirit and scope of the present invention.

Sequence listing

<110> university of Lonshler Physics (Rensselaer Polytechnic Institute)

<120> contraceptive vaccine based on sperm-associated protein CATSPER

<130> 104312-201

<140> PCT/US20/17449

<141> 2020-02-10

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<170> PatentIn version 3.5

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35 40 45

Val Gly His Pro Tyr Tyr Ser Ile Lys Lys Val Asn Lys Thr Val Val

50 55 60

Pro Lys Val Ser Gly Tyr Gln Tyr Arg Val Phe Lys Val Val Leu Pro

65 70 75 80

Asp Pro Asn Lys Phe Ala Leu Pro Asp Ser Ser Leu Phe Asp Pro Thr

85 90 95

Thr Gln Arg Leu Val Trp Ala Cys Thr Gly Leu Glu Val Gly Arg Gly

100 105 110

Gln Pro Leu Gly Val Gly Val Ser Gly His Pro Leu Leu Asn Lys Tyr

115 120 125

Asp Asp Val Glu Asn Ser Gly Gly Tyr Gly Cys Gly Pro Arg Glu Tyr

130 135 140

Ser Arg Ser Thr Ile Glu Gly Leu Glu Tyr Asn Gly Pro Cys Gly Asn

145 150 155 160

Pro Gly Gln Asp Asn Arg Val Asn Val Gly Met Asp Tyr Lys Gln Thr

165 170 175

Gln Leu Cys Met Val Gly Cys Ala Pro Pro Leu Gly Glu His Trp Gly

180 185 190

Lys Gly Thr Gln Cys Ser Asn Thr Ser Val Gln Asn Gly Asp Cys Pro

195 200 205

Pro Leu Glu Leu Ile Thr Ser Val Ile Gln Asp Gly Asp Met Val Asp

210 215 220

Thr Gly Phe Gly Ala Met Asn Phe Ala Asp Leu Gln Thr Asn Lys Ser

225 230 235 240

Asp Val Pro Leu Asp Ile Cys Gly Thr Val Cys Lys Tyr Pro Asp Tyr

245 250 255

Leu Gln Met Ala Ala Asp Pro Tyr Gly Asp Arg Leu Phe Phe Tyr Leu

260 265 270

Arg Lys Glu Gln Met Phe Ala Arg His Phe Phe Asn Arg Ala Gly Thr

275 280 285

Val Gly Glu Pro Val Pro Asp Asp Leu Leu Val Lys Gly Gly Asn Asn

290 295 300

Arg Ser Ser Val Ala Ser Ser Ile Tyr Val His Thr Pro Ser Gly Ser

305 310 315 320

Leu Val Ser Ser Glu Ala Gln Leu Phe Asn Lys Pro Tyr Trp Leu Gln

325 330 335

Lys Ala Gln Gly His Asn Asn Gly Ile Cys Trp Gly Asn His Leu Phe

340 345 350

Val Thr Val Val Asp Thr Thr Arg Ser Thr Asn Met Thr Leu Cys Ala

355 360 365

Ser Val Ser Lys Ser Ala Thr Tyr Thr Asn Ser Asp Tyr Lys Glu Tyr

370 375 380

Met Arg His Val Glu Glu Phe Asp Leu Gln Phe Ile Phe Gln Leu Cys

385 390 395 400

Ser Ile Thr Leu Ser Ala Glu Val Met Ala Tyr Ile His Thr Met Asn

405 410 415

Pro Ser Val Leu Glu Asp Trp Asn Phe Gly Leu Ser Pro Pro Pro Asn

420 425 430

Gly Thr Leu Glu Asp Thr Tyr Arg Tyr Val Gln Ser Gln Ala Ile Thr

435 440 445

Cys Gln Lys Pro Thr Pro Glu Lys Glu Lys Gln Asp Pro Tyr Lys Asp

450 455 460

Met Ser Phe Trp Glu Val Asn Leu Lys Glu Lys Phe Ser Ser Glu Leu

465 470 475 480

Asp Gln Phe Pro Leu Gly Arg Lys Phe Leu Leu Gln Ser Gly Tyr Arg

485 490 495

Gly Arg Thr Ser Ala Arg Thr Gly Ile

500 505

<210> 3

<211> 499

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 3

Ser Thr Met Glu Phe Met Trp Arg Pro Ser Asp Ser Thr Val Tyr Val

1 5 10 15

Pro Pro Pro Asn Pro Val Ser Lys Val Val Ala Thr Asp Ala Tyr Val

20 25 30

Lys Arg Thr Asn Ile Phe Tyr His Ala Ser Ser Ser Arg Leu Leu Ala

35 40 45

Val Gly His Pro Tyr Tyr Ser Ile Cys Gly Pro Asn Ser Leu Ser Tyr

50 55 60

Ser Phe Tyr Asn His Ser Leu Phe Arg Gly Pro Cys Val Val Pro Lys

65 70 75 80

Val Ser Gly Tyr Gln Tyr Arg Val Phe Lys Val Val Leu Pro Asp Pro

85 90 95

Asn Lys Phe Ala Leu Pro Asp Ser Ser Leu Phe Asp Pro Thr Thr Gln

100 105 110

Arg Leu Val Trp Ala Cys Thr Gly Leu Glu Val Gly Arg Gly Gln Pro

115 120 125

Leu Gly Val Gly Val Ser Gly His Pro Leu Leu Asn Lys Tyr Asp Asp

130 135 140

Val Glu Asn Ser Gly Gly Tyr Gly Gly Asn Pro Gly Gln Asp Asn Arg

145 150 155 160

Val Asn Val Gly Met Asp Tyr Lys Gln Thr Gln Leu Cys Met Val Gly

165 170 175

Cys Ala Pro Pro Leu Gly Glu His Trp Gly Lys Gly Thr Gln Cys Ser

180 185 190

Asn Thr Ser Val Gln Asn Gly Asp Cys Pro Pro Leu Glu Leu Ile Thr

195 200 205

Ser Val Ile Gln Asp Gly Asp Met Val Asp Thr Gly Phe Gly Ala Met

210 215 220

Asn Phe Ala Asp Leu Gln Thr Asn Lys Ser Asp Val Pro Leu Asp Ile

225 230 235 240

Cys Gly Thr Val Cys Lys Tyr Pro Asp Tyr Leu Gln Met Ala Ala Asp

245 250 255

Pro Tyr Gly Asp Arg Leu Phe Phe Tyr Leu Arg Lys Glu Gln Met Phe

260 265 270

Ala Arg His Phe Phe Asn Arg Ala Gly Thr Val Gly Glu Pro Val Pro

275 280 285

Asp Asp Leu Leu Val Lys Gly Gly Asn Asn Arg Ser Ser Val Ala Ser

290 295 300

Ser Ile Tyr Val His Thr Pro Ser Gly Ser Leu Val Ser Ser Glu Ala

305 310 315 320

Gln Leu Phe Asn Lys Pro Tyr Trp Leu Gln Lys Ala Gln Gly His Asn

325 330 335

Asn Gly Ile Cys Trp Gly Asn His Leu Phe Val Thr Val Val Asp Thr

340 345 350

Thr Arg Ser Thr Asn Met Thr Leu Cys Ala Ser Val Ser Lys Ser Ala

355 360 365

Thr Tyr Thr Asn Ser Asp Tyr Lys Glu Tyr Met Arg His Val Glu Glu

370 375 380

Phe Asp Leu Gln Phe Ile Phe Gln Leu Cys Ser Ile Thr Leu Ser Ala

385 390 395 400

Glu Val Met Ala Tyr Ile His Thr Met Asn Pro Ser Val Leu Glu Asp

405 410 415

Trp Asn Phe Gly Leu Ser Pro Pro Pro Asn Gly Thr Leu Glu Asp Thr

420 425 430

Tyr Arg Tyr Val Gln Ser Gln Ala Ile Thr Cys Gln Lys Pro Thr Pro

435 440 445

Glu Lys Glu Lys Gln Asp Pro Tyr Lys Asp Met Ser Phe Trp Glu Val

450 455 460

Asn Leu Lys Glu Lys Phe Ser Ser Glu Leu Asp Gln Phe Pro Leu Gly

465 470 475 480

Arg Lys Phe Leu Leu Gln Ser Gly Tyr Arg Gly Arg Thr Ser Ala Arg

485 490 495

Thr Gly Ile

<210> 4

<211> 499

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 4

Ser Thr Met Glu Phe Met Trp Arg Pro Ser Asp Ser Thr Val Tyr Val

1 5 10 15

Pro Pro Pro Asn Pro Val Ser Lys Val Val Ala Thr Asp Ala Tyr Val

20 25 30

Lys Arg Thr Asn Ile Phe Tyr His Ala Ser Ser Ser Arg Leu Leu Ala

35 40 45

Val Gly His Pro Tyr Tyr Ser Ile Cys Gly Pro Glu Ile Glu Leu Met

50 55 60

Glu Ser Thr Asn Thr Ala Leu Trp Pro Gly Pro Cys Val Val Pro Lys

65 70 75 80

Val Ser Gly Tyr Gln Tyr Arg Val Phe Lys Val Val Leu Pro Asp Pro

85 90 95

Asn Lys Phe Ala Leu Pro Asp Ser Ser Leu Phe Asp Pro Thr Thr Gln

100 105 110

Arg Leu Val Trp Ala Cys Thr Gly Leu Glu Val Gly Arg Gly Gln Pro

115 120 125

Leu Gly Val Gly Val Ser Gly His Pro Leu Leu Asn Lys Tyr Asp Asp

130 135 140

Val Glu Asn Ser Gly Gly Tyr Gly Gly Asn Pro Gly Gln Asp Asn Arg

145 150 155 160

Val Asn Val Gly Met Asp Tyr Lys Gln Thr Gln Leu Cys Met Val Gly

165 170 175

Cys Ala Pro Pro Leu Gly Glu His Trp Gly Lys Gly Thr Gln Cys Ser

180 185 190

Asn Thr Ser Val Gln Asn Gly Asp Cys Pro Pro Leu Glu Leu Ile Thr

195 200 205

Ser Val Ile Gln Asp Gly Asp Met Val Asp Thr Gly Phe Gly Ala Met

210 215 220

Asn Phe Ala Asp Leu Gln Thr Asn Lys Ser Asp Val Pro Leu Asp Ile

225 230 235 240

Cys Gly Thr Val Cys Lys Tyr Pro Asp Tyr Leu Gln Met Ala Ala Asp

245 250 255

Pro Tyr Gly Asp Arg Leu Phe Phe Tyr Leu Arg Lys Glu Gln Met Phe

260 265 270

Ala Arg His Phe Phe Asn Arg Ala Gly Thr Val Gly Glu Pro Val Pro

275 280 285

Asp Asp Leu Leu Val Lys Gly Gly Asn Asn Arg Ser Ser Val Ala Ser

290 295 300

Ser Ile Tyr Val His Thr Pro Ser Gly Ser Leu Val Ser Ser Glu Ala

305 310 315 320

Gln Leu Phe Asn Lys Pro Tyr Trp Leu Gln Lys Ala Gln Gly His Asn

325 330 335

Asn Gly Ile Cys Trp Gly Asn His Leu Phe Val Thr Val Val Asp Thr

340 345 350

Thr Arg Ser Thr Asn Met Thr Leu Cys Ala Ser Val Ser Lys Ser Ala

355 360 365

Thr Tyr Thr Asn Ser Asp Tyr Lys Glu Tyr Met Arg His Val Glu Glu

370 375 380

Phe Asp Leu Gln Phe Ile Phe Gln Leu Cys Ser Ile Thr Leu Ser Ala

385 390 395 400

Glu Val Met Ala Tyr Ile His Thr Met Asn Pro Ser Val Leu Glu Asp

405 410 415

Trp Asn Phe Gly Leu Ser Pro Pro Pro Asn Gly Thr Leu Glu Asp Thr

420 425 430

Tyr Arg Tyr Val Gln Ser Gln Ala Ile Thr Cys Gln Lys Pro Thr Pro

435 440 445

Glu Lys Glu Lys Gln Asp Pro Tyr Lys Asp Met Ser Phe Trp Glu Val

450 455 460

Asn Leu Lys Glu Lys Phe Ser Ser Glu Leu Asp Gln Phe Pro Leu Gly

465 470 475 480

Arg Lys Phe Leu Leu Gln Ser Gly Tyr Arg Gly Arg Thr Ser Ala Arg

485 490 495

Thr Gly Ile

<210> 5

<211> 497

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 5

Ser Thr Met Glu Phe Met Trp Arg Pro Ser Asp Ser Thr Val Tyr Val

1 5 10 15

Pro Pro Pro Asn Pro Val Ser Lys Val Val Ala Thr Asp Ala Tyr Val

20 25 30

Lys Arg Thr Asn Ile Phe Tyr His Ala Ser Ser Ser Arg Leu Leu Ala

35 40 45

Val Gly His Pro Tyr Tyr Ser Ile Cys Gly Pro Gly Thr Asn Tyr Asp

50 55 60

Ile Gln Phe Glu Phe Phe Arg Gly Pro Cys Val Val Pro Lys Val Ser

65 70 75 80

Gly Tyr Gln Tyr Arg Val Phe Lys Val Val Leu Pro Asp Pro Asn Lys

85 90 95

Phe Ala Leu Pro Asp Ser Ser Leu Phe Asp Pro Thr Thr Gln Arg Leu

100 105 110

Val Trp Ala Cys Thr Gly Leu Glu Val Gly Arg Gly Gln Pro Leu Gly

115 120 125

Val Gly Val Ser Gly His Pro Leu Leu Asn Lys Tyr Asp Asp Val Glu

130 135 140

Asn Ser Gly Gly Tyr Gly Gly Asn Pro Gly Gln Asp Asn Arg Val Asn

145 150 155 160

Val Gly Met Asp Tyr Lys Gln Thr Gln Leu Cys Met Val Gly Cys Ala

165 170 175

Pro Pro Leu Gly Glu His Trp Gly Lys Gly Thr Gln Cys Ser Asn Thr

180 185 190

Ser Val Gln Asn Gly Asp Cys Pro Pro Leu Glu Leu Ile Thr Ser Val

195 200 205

Ile Gln Asp Gly Asp Met Val Asp Thr Gly Phe Gly Ala Met Asn Phe

210 215 220

Ala Asp Leu Gln Thr Asn Lys Ser Asp Val Pro Leu Asp Ile Cys Gly

225 230 235 240

Thr Val Cys Lys Tyr Pro Asp Tyr Leu Gln Met Ala Ala Asp Pro Tyr

245 250 255

Gly Asp Arg Leu Phe Phe Tyr Leu Arg Lys Glu Gln Met Phe Ala Arg

260 265 270

His Phe Phe Asn Arg Ala Gly Thr Val Gly Glu Pro Val Pro Asp Asp

275 280 285

Leu Leu Val Lys Gly Gly Asn Asn Arg Ser Ser Val Ala Ser Ser Ile

290 295 300

Tyr Val His Thr Pro Ser Gly Ser Leu Val Ser Ser Glu Ala Gln Leu

305 310 315 320

Phe Asn Lys Pro Tyr Trp Leu Gln Lys Ala Gln Gly His Asn Asn Gly

325 330 335

Ile Cys Trp Gly Asn His Leu Phe Val Thr Val Val Asp Thr Thr Arg

340 345 350

Ser Thr Asn Met Thr Leu Cys Ala Ser Val Ser Lys Ser Ala Thr Tyr

355 360 365

Thr Asn Ser Asp Tyr Lys Glu Tyr Met Arg His Val Glu Glu Phe Asp

370 375 380

Leu Gln Phe Ile Phe Gln Leu Cys Ser Ile Thr Leu Ser Ala Glu Val

385 390 395 400

Met Ala Tyr Ile His Thr Met Asn Pro Ser Val Leu Glu Asp Trp Asn

405 410 415

Phe Gly Leu Ser Pro Pro Pro Asn Gly Thr Leu Glu Asp Thr Tyr Arg

420 425 430

Tyr Val Gln Ser Gln Ala Ile Thr Cys Gln Lys Pro Thr Pro Glu Lys

435 440 445

Glu Lys Gln Asp Pro Tyr Lys Asp Met Ser Phe Trp Glu Val Asn Leu

450 455 460

Lys Glu Lys Phe Ser Ser Glu Leu Asp Gln Phe Pro Leu Gly Arg Lys

465 470 475 480

Phe Leu Leu Gln Ser Gly Tyr Arg Gly Arg Thr Ser Ala Arg Thr Gly

485 490 495

Ile

<210> 6

<211> 501

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 6

Ser Thr Met Glu Phe Met Trp Arg Pro Ser Asp Ser Thr Val Tyr Val

1 5 10 15

Pro Pro Pro Asn Pro Val Ser Lys Val Val Ala Thr Asp Ala Tyr Val

20 25 30

Lys Arg Thr Asn Ile Phe Tyr His Ala Ser Ser Ser Arg Leu Leu Ala

35 40 45

Val Gly His Pro Tyr Tyr Ser Ile Lys Lys Val Asn Lys Thr Val Val

50 55 60

Pro Lys Val Ser Gly Tyr Gln Tyr Arg Val Phe Lys Val Val Leu Pro

65 70 75 80

Asp Pro Asn Lys Phe Ala Leu Pro Asp Ser Ser Leu Phe Asp Pro Thr

85 90 95

Thr Gln Arg Leu Val Trp Ala Cys Thr Gly Leu Glu Val Gly Arg Gly

100 105 110

Gln Pro Leu Gly Val Gly Val Ser Gly His Pro Leu Leu Asn Lys Tyr

115 120 125

Asp Asp Val Glu Asn Ser Gly Gly Tyr Gly Gly Asn Pro Gly Gln Asp

130 135 140

Asn Arg Val Asn Val Gly Met Asp Tyr Lys Gln Thr Gln Leu Cys Met

145 150 155 160

Val Gly Cys Ala Pro Pro Leu Gly Glu His Trp Gly Lys Gly Thr Gln

165 170 175

Cys Ser Asn Thr Ser Val Gln Asn Gly Asp Cys Pro Pro Leu Glu Leu

180 185 190

Ile Thr Ser Val Ile Gln Asp Gly Asp Met Val Asp Thr Gly Phe Gly

195 200 205

Ala Met Asn Phe Ala Asp Leu Gln Thr Asn Lys Ser Asp Val Pro Leu

210 215 220

Asp Ile Cys Gly Thr Val Cys Lys Tyr Pro Asp Tyr Leu Gln Met Ala

225 230 235 240

Ala Asp Pro Tyr Gly Asp Arg Leu Phe Phe Tyr Leu Arg Lys Glu Gln

245 250 255

Met Phe Ala Arg His Phe Phe Asn Arg Ala Gly Thr Val Gly Glu Pro

260 265 270

Val Pro Asp Asp Leu Leu Val Lys Gly Gly Asn Asn Arg Ser Ser Val

275 280 285

Ala Ser Ser Ile Tyr Val His Thr Pro Ser Gly Ser Leu Val Ser Ser

290 295 300

Glu Ala Gln Leu Phe Asn Lys Pro Tyr Trp Leu Gln Lys Ala Gln Gly

305 310 315 320

His Asn Asn Gly Ile Cys Trp Gly Asn His Leu Phe Val Thr Val Val

325 330 335

Asp Thr Thr Arg Ser Thr Asn Met Thr Leu Cys Ala Ser Val Cys Gly

340 345 350

Pro Arg Ala Leu Phe Gln Asp Ser Asp Pro Lys Arg Phe Gln Asn Gly

355 360 365

Pro Cys Thr Tyr Thr Asn Ser Asp Tyr Lys Glu Tyr Met Arg His Val

370 375 380

Glu Glu Phe Asp Leu Gln Phe Ile Phe Gln Leu Cys Ser Ile Thr Leu

385 390 395 400

Ser Ala Glu Val Met Ala Tyr Ile His Thr Met Asn Pro Ser Val Leu

405 410 415

Glu Asp Trp Asn Phe Gly Leu Ser Pro Pro Pro Asn Gly Thr Leu Glu

420 425 430

Asp Thr Tyr Arg Tyr Val Gln Ser Gln Ala Ile Thr Cys Gln Lys Pro

435 440 445

Thr Pro Glu Lys Glu Lys Gln Asp Pro Tyr Lys Asp Met Ser Phe Trp

450 455 460

Glu Val Asn Leu Lys Glu Lys Phe Ser Ser Glu Leu Asp Gln Phe Pro

465 470 475 480

Leu Gly Arg Lys Phe Leu Leu Gln Ser Gly Tyr Arg Gly Arg Thr Ser

485 490 495

Ala Arg Thr Gly Ile

500

<210> 7

<211> 498

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 7

Ser Thr Met Glu Phe Met Trp Arg Pro Ser Asp Ser Thr Val Tyr Val

1 5 10 15

Pro Pro Pro Asn Pro Val Ser Lys Val Val Ala Thr Asp Ala Tyr Val

20 25 30

Lys Arg Thr Asn Ile Phe Tyr His Ala Ser Ser Ser Arg Leu Leu Ala

35 40 45

Val Gly His Pro Tyr Tyr Ser Ile Lys Lys Val Asn Lys Thr Val Val

50 55 60

Pro Lys Val Ser Gly Tyr Gln Tyr Arg Val Phe Lys Val Val Leu Pro

65 70 75 80

Asp Pro Asn Lys Phe Ala Leu Pro Asp Ser Ser Leu Phe Asp Pro Thr

85 90 95

Thr Gln Arg Leu Val Trp Ala Cys Thr Gly Leu Glu Val Gly Arg Gly

100 105 110

Gln Pro Leu Gly Val Gly Val Ser Gly His Pro Leu Leu Asn Lys Tyr

115 120 125

Asp Asp Val Glu Asn Ser Gly Gly Tyr Gly Gly Asn Pro Gly Gln Asp

130 135 140

Asn Arg Val Asn Val Gly Met Asp Tyr Lys Gln Thr Gln Leu Cys Met

145 150 155 160

Val Gly Cys Ala Pro Pro Leu Gly Glu His Trp Gly Lys Gly Thr Gln

165 170 175

Cys Ser Asn Thr Ser Val Gln Asn Gly Asp Cys Pro Pro Leu Glu Leu

180 185 190

Ile Thr Ser Val Ile Gln Asp Gly Asp Met Val Asp Thr Gly Phe Gly

195 200 205

Ala Met Asn Phe Ala Asp Leu Gln Thr Asn Lys Ser Asp Val Pro Leu

210 215 220

Asp Ile Cys Gly Thr Val Cys Lys Tyr Pro Asp Tyr Leu Gln Met Ala

225 230 235 240

Ala Asp Pro Tyr Gly Asp Arg Leu Phe Phe Tyr Leu Arg Lys Glu Gln

245 250 255

Met Phe Ala Arg His Phe Phe Asn Arg Ala Gly Thr Val Gly Glu Pro

260 265 270

Val Pro Asp Asp Leu Leu Val Lys Gly Gly Asn Asn Arg Ser Ser Val

275 280 285

Ala Ser Ser Ile Tyr Val His Thr Pro Ser Gly Ser Leu Val Ser Ser

290 295 300

Glu Ala Gln Leu Phe Asn Lys Pro Tyr Trp Leu Gln Lys Ala Gln Gly

305 310 315 320

His Asn Asn Gly Ile Cys Trp Gly Asn His Leu Phe Val Thr Val Val

325 330 335

Asp Thr Thr Arg Ser Thr Asn Met Thr Leu Cys Ala Ser Val Cys Gly

340 345 350

Pro Gln Asp Ile Trp Lys Val Pro Glu Ser Ser Arg Gly Pro Cys Thr

355 360 365

Tyr Thr Asn Ser Asp Tyr Lys Glu Tyr Met Arg His Val Glu Glu Phe

370 375 380

Asp Leu Gln Phe Ile Phe Gln Leu Cys Ser Ile Thr Leu Ser Ala Glu

385 390 395 400

Val Met Ala Tyr Ile His Thr Met Asn Pro Ser Val Leu Glu Asp Trp

405 410 415

Asn Phe Gly Leu Ser Pro Pro Pro Asn Gly Thr Leu Glu Asp Thr Tyr

420 425 430

Arg Tyr Val Gln Ser Gln Ala Ile Thr Cys Gln Lys Pro Thr Pro Glu

435 440 445

Lys Glu Lys Gln Asp Pro Tyr Lys Asp Met Ser Phe Trp Glu Val Asn

450 455 460

Leu Lys Glu Lys Phe Ser Ser Glu Leu Asp Gln Phe Pro Leu Gly Arg

465 470 475 480

Lys Phe Leu Leu Gln Ser Gly Tyr Arg Gly Arg Thr Ser Ala Arg Thr

485 490 495

Gly Ile

<210> 8

<211> 12

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 8

Gln Thr Phe Thr Glu Leu Glu Ile Arg Gly Glu Trp

1 5 10

<210> 9

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 9

Asn Ser Leu Ser Tyr Ser Phe Tyr Asn His Ser Leu Phe Arg

1 5 10

<210> 10

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 10

Arg Ala Leu Phe Gln Asp Ser Asp Pro Lys Arg Phe Gln Asn

1 5 10

<210> 11

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 11

Glu Ile Glu Leu Met Glu Ser Thr Asn Thr Ala Leu Trp Pro

1 5 10

<210> 12

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 12

Phe Arg Glu Tyr Ser Arg Ser Thr Ile Glu Gly Leu Glu Tyr

1 5 10

<210> 13

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 13

Arg Glu Tyr Ser Arg Ser Thr Ile Glu Gly Leu Glu Tyr Asn

1 5 10

<210> 14

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 14

Gln Asp Ile Trp Lys Val Pro Glu Ser Ser Arg

1 5 10

<210> 15

<211> 12

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 15

Gly Thr Asn Tyr Asp Ile Gln Phe Glu Phe Phe Arg

1 5 10

<210> 16

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 16

Arg Lys Ile Lys Gly Asn His Ser Ala Tyr Leu His Phe Ala Asp Gly

1 5 10 15

<210> 17

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 17

Gly Val Thr Asp Arg Gly Asp Leu Glu Asn Trp Gly Asn

1 5 10

<210> 18

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 18

Gln Glu Glu Leu Asp Lys Arg Lys Phe Thr Val Ser Arg

1 5 10

<210> 19

<211> 12

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 19

Arg Thr Asn Ser Tyr Leu Gly Gln Lys His Tyr Glu

1 5 10

<210> 20

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 20

Tyr Thr Asp Phe Gln Met Asp Glu Arg Glu Tyr Ala Met Glu

1 5 10

<210> 21

<211> 20

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 21

Arg Arg Val Lys Asp Gln Asn Arg Gly Lys Val Arg Val Ala Gln Lys

1 5 10 15

His Pro Glu Thr

20

<210> 22

<211> 18

<212> PRT

<213> Artificial sequence

<220>

<223> description of artificial sequences: synthetic peptides

<400> 22

Asp Gly Thr Val Tyr Leu Arg Thr Glu Asp Glu Phe Thr Lys Leu Asp

1 5 10 15

Glu Ser