Superparamagnetic and highly porous polymer particles for diagnostic applications
阅读说明：本技术 用于诊断应用的超顺磁和高度多孔聚合物颗粒 (Superparamagnetic and highly porous polymer particles for diagnostic applications ) 是由 J.C.博莱 M.E.西尔韦斯特雷 S.胡格 C.拜尔达 于 2018-04-12 设计创作，主要内容包括：本发明涉及磁性颗粒,其中每个颗粒包含聚合物基质和至少一个磁芯(M),其中所述聚合物基质包含至少一种交联的聚合物,且其中所述磁性颗粒具有1-60微米的范围内的粒径,进一步,本发明涉及制备这样的颗粒的方法,和通过所述方法可获得的或获得的颗粒。而且,本发明涉及这些磁性颗粒用于定性和/或定量测定流体中的至少一种分析物的用途。进一步,本发明涉及用于测定流体样品中的至少一种分析物的方法,其包括使本发明的磁性颗粒或通过本发明的方法获得的磁性颗粒与包含或怀疑包含至少一种分析物的流体样品接触。(The present invention relates to magnetic particles, wherein each particle comprises a polymer matrix and at least magnetic cores (M), wherein the polymer matrix comprises at least cross-linked polymers, and wherein the magnetic particles have a particle size in the range of 1-60 microns, step , to a method of preparing such particles, and to particles obtainable or obtained by said method, furthermore, the present invention relates to the use of these magnetic particles for the qualitative and/or quantitative determination of at least analytes in a fluid, step , the present invention relates to a method for the determination of at least analytes in a fluid sample, comprising contacting a magnetic particle of the present invention or a magnetic particle obtained by a method of the present invention with a fluid sample comprising or suspected to comprise at least analytes.)
1. Magnetic particle comprising a polymer matrix (P) and at least magnetic cores (M), preferably at least two magnetic cores, wherein the polymer matrix comprises a hypercrosslinked polymer and wherein the magnetic particle has a particle size in the range of 5-40 microns as determined according to ISO 13320.
2. The magnetic particle according to claim 1, wherein the polymer matrix comprises pores having a pore size of less than 100 nm, preferably less than or equal to 50 nm, as determined according to ISO 15901-3.
3. The magnetic particle according to claim 2, wherein at least 90% of all pores present in the polymer matrix (P) have a pore size of less than 10 nm and at least 50% of all pores present in the polymer matrix have a pore size of less than 5 nm, as determined according to ISO 15901-3, preferably wherein the polymer matrix (P) does not comprise macropores having a pore size of more than 50 nm.
4. A magnetic particle as claimed in claim 2 or 3 wherein the particle has a size of 50 to 2500 m2BET specific surface area in the range of/g, as determined according to ISO 9277.
5. The magnetic particles of any of claims 1 to 4, wherein the magnetic particles are magnetic particlesThe magnetic particles have a magnetic particle size of at least 1A m2/kg, preferably at least 10A m2Saturation magnetization of/kg.
6. The magnetic particles of any of claims 1 to 5, wherein the magnetic particles are superparamagnetic.
7. The magnetic particle of any of claims 1 to 6 wherein the at least magnetic cores (M) comprise at least magnetic nanoparticles, preferably at least iron oxide nanoparticles, more preferably Fe3O4-nanoparticles.
8. A magnetic particle as claimed in claim 7 wherein said magnetic core (M) comprises, more preferably consists of, at least nanoparticles and a coating C1.
9. The magnetic particles of of claims 1 to 6 wherein the at least magnetic cores (M) comprise, preferably consist of, super particles and optionally comprise a coating of C1.
10. The magnetic particle of claim 9 wherein the nanoparticle is comprised of aggregated nanoparticles.
11. The magnetic particle according to claim 9 or 10, wherein the nanoparticle consists of aggregated nanoparticles, and preferably consists of more than 20 aggregated nanoparticles, and more preferably consists of 100-150 ten thousand nanoparticles.
12. The magnetic particle of of claims 9 to 11, wherein the at least magnetic cores (M) comprise, preferably consist of, a super particle and at least coating layers C2, the coating layers preferably being deposited on the surface of the nanoparticle,
and wherein the coating C2 is preferably selected from the group consisting of dicarboxylic acids, dicarboxylic acid salts, dicarboxylic acid derivatives, polyacrylic acid salts, polyacrylic acid derivatives, tricarboxylic acids, tricarboxylic acid salts, tricarboxylic acid derivatives, amino acids, amino acid salts, amino acid derivatives, surfactants, salts of surfactants, fatty acids, fatty acid salts and fatty acid derivatives.
13. The magnetic particles of any of of claims 7 to 12 wherein the at least coating C1 is selected from the group consisting of surfactants, silica, silicates, silanes, phosphates, phosphonates, phosphonic acids, and mixtures of two or more thereof.
14. The magnetic particles of claims 1 to 13 wherein the polymer matrix (P) comprises a copolymer obtained or obtainable by a process comprising copolymerizing suitable monomer building blocks, which are cross-linkers, in the presence of at least monomer building blocks, wherein preferably 5 to 90vol% of all monomer building blocks are cross-linkers, more preferably divinylbenzene.
15. A method for the preparation of a magnetic particle comprising a polymer matrix (P) and at least magnetic cores (M), preferably at least two magnetic cores (M), wherein the polymer matrix (P) comprises at least crosslinked polymers, wherein the magnetic particle has a particle size in the range of 5-40 microns as determined according to ISO13320, the method comprising:
(i) providing at least magnetic cores (M), preferably at least two magnetic cores (M),
(ii) providing a precursor molecule of the polymer,
(iii) (iii) polymerizing the polymer precursor molecules according to (ii) in the presence of at least magnetic cores (M) thereby forming particles comprising at least magnetic cores (M), preferably at least two magnetic cores (M), which are embedded in a polymer matrix (P1), wherein the polymer matrix (P1) preferably comprises, more preferably consists of, a cross-linked polymer, and
(iv) (iv) hypercrosslinking the polymer matrix (P1) of the polymer particles obtained in (iii),
to obtain magnetic particles.
16. The method of claim 15, wherein the hypercrosslinking in (iv) is performed by a friedel-crafts reaction.
17. Magnetic particles obtained or obtainable by the method of claim 15 or 16.
18. Use of the magnetic particles of any of claims 1-14 or the magnetic particles of claim 17 for qualitative and/or quantitative determination of at least analytes in a fluid.
19. The use of claim 18, wherein the analyte is selected from the group consisting of steroids, vitamins, drugs, sugars, organic compounds, proteins, nucleic acids, sugars, and mixtures of two or more thereof.
20. A method for determining at least analytes in a fluid sample, comprising the steps of:
a) contacting the magnetic particles of any of claims 1 to 14 or claim 17 with a fluid sample containing or suspected of containing at least analytes, and
b) determining at least analytes bound to the magnetic particles.
Technical Field
The present invention relates to magnetic particles, wherein each particle comprises a polymer matrix (P) and at least magnetic cores (M), wherein the polymer matrix comprises at least cross-linked polymers, and wherein the magnetic particles have a particle size in the range of 1-60 microns, step , the present invention relates to a method of preparing such particles, and to particles obtainable or obtained by the method, furthermore, the present invention relates to the use of these magnetic particles for the qualitative and/or quantitative determination of at least analytes in a fluid, step , the present invention relates to a method for the determination of at least analytes in a fluid sample, comprising contacting the magnetic particles of the present invention or the magnetic particles obtained by the method of the present invention with a fluid sample comprising or suspected to comprise at least analytes.
RELATED ART
Magnetic properties are of great interest here, because they only show a magnetization when an external magnetic field is applied, the magnetization assumes zero (no "memory effect") in the absence of an external magnetic field, e.g. EP2003455a1 and EP2015074a1 describe the extraction of analytes from human samples by using magnetic particles on LC/MS systems.
A high specific surface area on the magnetic particles is required to enrich the analyte from the human sample. In order to increase the surface area to more than 1000 m2 disadvantages are that the high density of silica and titanium leads to a decrease in magnetization with increasing film thickness, furthermore, by using silica or titanium oxide, only systems with mesopores (pores >2 nm) can be developed, but especially for small analytes, materials with micropores (pores < 2 nm) are preferred.
key requirements for particle automation in enrichment-workflow-MS technology are for high throughput rapid magnetic separation (< 5 s), particle size and saturation magnetization are key properties, therefore, there is a need to have high saturation magnetization (> 5 Am)2kg-1) And large (> 2 μm) size particles. In addition, carryover of particles must be avoided for robustness of the system. Therefore, the particles need to have high magnetization and a particle diameter of more than 1 μm.
Microporous particles with a polymer matrix, e.g. in Yang et alPolym. Chem.2013, 4, 1425. According to Yang et al, iron oxide nanoparticles are first coated with oleic acid and then embedded in a polystyrene matrix by microemulsion polymerization. To achieve a high specific surface area, the nanoparticles are finally hypercrosslinked by a Friedel-crafts reaction, using FeCl3As catalyst dimethoxymethane was used as porosity-dependent crosslinker. The resulting particles had an average size of 100 nm and 4.1A m2kg-1The saturation magnetization of (2). The particles are described as useful for extracting organic molecules from water and also as drug carriers to control the delivery of ibuprofen drugs, however, they only show relatively small particle size and low saturation magnetization, which makes them non-viableIs beneficial to the automation process.
Xu et al describe the synthesis of citrate-stabilized iron oxide nanoparticles having a particle size of about 300 nm (sPolym.Chem.2015, 6, 2892) these citrate stabilized nanoparticles were coated with 3- (trimethoxysilyl) propyl methacrylate (MEMO) and covered by a polystyrene shell by soap-free emulsion polymerization with this technique nanoparticles were embedded in the polymer particles in a second polymerization another layer was formed on the particlesSeed of a plant Swelling of seedsAnd as a final step by reaction with FeCl as a pore-forming catalyst3The hypercrosslinking reaction forms pores. However, in this way, only a size of about 400nm, 14.1A m, is achieved2kg-1The saturation magnetization of (2).
Thus, there remains a need for advantageous particles, particularly particles useful for automated processes.
Summary of The Invention
It is therefore an object of the present invention to provide porous magnetic particles having a relatively large particle diameter and a relatively high saturation magnetization. This problem is solved by the invention with the features of the independent patent claims. Advantageous embodiments of the invention which can be realized individually or in combination are given in the dependent claims and/or in the description and detailed embodiments below.
Thus, the terms "having," "including," or any grammatical variations thereof, may refer to the absence of other features in the entities described in this context, and the presence of or more other features, in addition to the features introduced by such terms.
Furthermore, it should be noted that the terms "at least /s", "/s" or similar expressions indicating that features or elements may be present times or more than times will generally only be used times when introducing the respective features or elements, hereinafter, in most cases, when referring to the respective features or elements, the expressions "at least /s" or "/s" will not be repeated, despite the fact that the respective features or elements may be present times or more than times.
Thus, the invention may be carried out by using alternative features as will be recognized by those skilled in the art, similarly, features introduced by "in the embodiments of the invention" or similar expressions are intended to be optional features without any limitation on the scope of alternative embodiments of the invention, and without any limitation on the scope of the invention, and the possibility of combinations of features introduced in this manner with other optional or non-optional features of the invention is not limited.
In an th aspect of the invention, there is provided a magnetic particle comprising a polymer matrix (P) and at least magnetic cores (M), wherein the polymer matrix comprises at least crosslinked polymers, and wherein the magnetic particle has a particle size in the range of 1-60 microns as determined according to ISO 13320.
In further steps, the invention relates to a method of preparing a magnetic particle comprising a polymer matrix (P) and at least magnetic cores (M), wherein the polymer matrix (P) comprises at least crosslinked polymers, wherein the magnetic particle has a particle size in the range of 1-60 microns, as determined according to ISO13320, the method comprising:
(i) providing at least magnetic cores (M), preferably at least two magnetic cores (M),
(ii) providing a precursor molecule of the polymer,
(iii) (iii) polymerizing the polymer precursor molecules according to (ii) in the presence of at least magnetic cores (M) thereby forming particles comprising at least magnetic cores (M), preferably at least two magnetic cores (M), embedded in a polymer matrix (P1), and
(iv) (iv) hypercrosslinking the polymer matrix (P1) of the polymer particles obtained in (iii),
to obtain magnetic particles.
In a third aspect, the present invention relates to the use of the magnetic particles described above and below or the magnetic particles obtainable by the methods described above and below for qualitative and/or quantitative determination of at least analytes in a fluid.
Furthermore, the present invention relates to a method for determining at least analytes in a fluid sample, comprising contacting a magnetic particle of the invention or a magnetic particle obtained by a method of the invention with a fluid sample comprising or suspected to comprise at least analytes.
Magnetic particles
The magnetic particles according to the present invention have a particle size of 1-60 micrometer, as determined according to ISO13320, more preferably the particle size is 5-55 micrometer, more preferably 10-50 micrometer, more preferably 15-45 micrometer, more preferably 20-40 micrometer, and especially 20-35 micrometer according to preferred embodiments the magnetic particles according to the present invention have a particle size of 5-40 micrometer, as determined according to ISO 13320.
As mentioned above, the magnetic particles according to the invention comprise a polymer matrix (P) and at least magnetic cores (M) according to preferred embodiments of the invention, said magnetic particles comprise more than magnetic cores (M), i.e. each particle preferably comprises at least , and preferably at least two magnetic cores (M) the magnetic core (M) comprises or more magnetic nanoparticles, such as 1-20 magnetic nanoparticles, preferably 1-10, more preferably 1-5, and most preferably 1-3 magnetic nanoparticles, or it may comprise more than 20 nanoparticles and, preferably 100-.
The invention therefore also relates to magnetic particles as described above, and to magnetic particles obtained or obtainable by the above-described process, wherein the particles comprise at least two magnetic cores (M) according to particularly preferred embodiments, the magnetic particles and the magnetic particles obtained or obtainable by the above-described process consist of at least two magnetic cores (M) and a polymer matrix (P).
Preferably, the amount of magnetic core (M) is chosen such that the desired saturation magnetization of the final particles is achieved. Preferably, the magnetic particles according to the invention, or the particles obtained or obtainable by the above-described process, have a particle size of at least 1A m2Saturation magnetization of/kg. Preferably, the saturation magnetization is at least 1A m2/kg, more preferably at least 2A m2/kg, more preferably at least 3A m2/kg, more preferably at least 4A m2/kg, more preferably at least 5A m2/kg, more preferably at least 6A m2/kg, more preferably at least 7A m2/kg, more preferably at least 8A m2/kg, more preferably at least 9A m2/kg, in particular at least 10A m2Kg, e.g. 10A m2/kg-20 A m2/kg, more preferably 10A m2/kg-30 A m2Kg, as determined according to ASTM A894/A894M.
As used herein, the term "substantially spherical" refers to particles having a rounded shape, which are preferably faceless or substantially free of sharp corners in certain embodiments, the substantially spherical particles generally have an average aspect ratio of less than 3:1 or 2:1, e.g., an aspect ratio of less than 1.5:1 or less than 1.2:1 in certain embodiments, the substantially spherical particles may have an aspect ratio of about 1:1R) Defined as the maximum diameter (d)max) And a minimum diameter (d) orthogonal theretomin) Function of (A)R= dmin/dmax). The diameter is determined by SEM or optical microscopy measurements.
As aboveThe BET specific surface area of the magnetic particles and the magnetic particles obtained or obtainable by the above method is preferably 50 to 2500 m2(iv)/g as determined according to ISO 9277. More preferably, the BET specific surface area of the magnetic particles is 100-1500 m2A/g, in particular of 300-2/g。
According to preferred embodiments of the present invention, the magnetic particles as described above, as well as the magnetic particles obtained or obtainable by the above-described methods, are superparamagnetic the term "superparamagnetic" is known to the skilled person and refers to the magnetic properties encountered in particular for particles smaller than a single magnetic monodomain such particles are stably oriented upon application of an external magnetic field until reaching a maximum of the overall magnetization, called the saturation magnetization, they relax upon removal of the magnetic field without hysteresis (no remanence) at room temperature.
Magnetic core (M)
As mentioned above, the magnetic particles according to the invention comprise at least magnetic cores (M) and preferably at least two magnetic cores (M), preferably the at least magnetic cores (M) comprise a compound selected from the group consisting of metals, metal carbides, metal nitrides, metal sulfides, metal phosphides, metal oxides, metal chelates and mixtures of two or more thereof the at least magnetic cores (M) may further comprise an alloy with a metal such as gold, silver, platinum or copper.
It is to be understood that each magnetic core (M) may comprise a mixture of two or more of the above groups, i.e. two or more of a metal, a metal carbide, a metal nitride, a metal sulfide, a metal phosphide, a metal oxide, a metal chelate and a mixture of two or more thereof. Furthermore, mixtures of two or more different metals, two or more different metal oxides, two or more different metal carbides, two or more different metal nitrides, two or more different metal sulfides, two or more different metal phosphides, two or more different metal chelates are conceivable.
Furthermore, it should be understood that in case the magnetic particles according to the present invention comprise more than magnetic cores (M), each magnetic core (M) present in a single particle may be identical or may be different from each other preferably all magnetic cores (M) comprised in magnetic particles are identical.
More preferably, said at least magnetic cores (M) comprise a metal oxide or a metal carbide.
In preferred embodiments, the at least magnetic cores (M) comprise a metal, metal carbide, metal nitride, metal sulfide, metal phosphide, metal oxide or metal chelate comprising at least transition metals preferred transition metals according to the invention include, but are not limited to, chromium, manganese, iron, cobalt, nickel, zinc, cadmium, nickel, gadolinium, copper and molybdenum more preferably the metal, metal carbide, metal nitride, metal sulfide, metal phosphide, metal oxide or metal chelate comprises at least iron more preferably the at least magnetic cores (M) comprise an iron oxide, in particular an iron oxide selected from the group consisting of Fe3O4、α-Fe2O3、γ-Fe2O3、MnFexOy、CoFexOy、NiFexOy、CuFexOy、ZnFexOy、CdFexOy、BaFexO and SrFexO, wherein x and y vary depending on the synthesis method, and wherein x is preferably an integer from 1 to 3, more preferably 2, and wherein y is preferably 3 or 4, most preferably, the at least magnetic cores (M) comprise Fe3O4。
Thus, the present invention also relates to magnetic particles as described above, and to magnetic particles obtained or obtainable by the above method, wherein said at least magnetic cores (M) comprise iron oxide, most preferably said at least magnetic cores (M) comprise Fe3O4。
The magnetic core (M) preferably comprises, more preferably consists of, nanoparticles and a coating C1.
Nanoparticles
The nanoparticles are preferably the part of the particles that exhibits magnetism, preferably superparamagnetism. Nanoparticles are also sometimes referred to herein as "magnetic nanoparticles".
Preferably, the at least nanoparticles comprise, preferably consist of, at least magnetic, preferably superparamagnetic, nanoparticles and optionally coatings, e.g. coating C2.
The term "nanoparticles" as used herein refers to particles smaller than 100 nm in at least dimensions, i.e. having a diameter of less than 100 nm preferably the nanoparticles according to the present invention have a diameter of 1-20 nm, preferably 4-15 nm, determined according to TEM measurements thus, according to preferred embodiments, the present invention also relates to magnetic particles as described above, as well as to magnetic particles obtained or obtainable by the above method, wherein the magnetic particles comprise at least magnetic cores (M) comprising at least nanoparticles and optionally coatings, e.g. coating C2.
Preferably each nanoparticle has a diameter of 1-20 nm, preferably 4-15 nm, as determined according to TEM measurements preferably said at least magnetic nanoparticles are superparamagnetic.
The magnetic core (M) may comprise only nanoparticles or more than nanoparticles, in embodiments it comprises 1-20 nanoparticles, in another embodiments it comprises 100-150 ten thousand nanoparticles, more preferably 750-750,000 nanoparticles, more preferably 1,750-320,000 nanoparticles, especially 90,000-320,000 nanoparticles, the nanoparticles may be present as individual (i.e. separate) particles as magnetic cores or they may be aggregates consisting of several nanoparticles, depending on the number of nanoparticles comprised, these aggregates may have different sizes.
Comprises 1-20 nanoparticlesMagnetic core of particle (M)
According to the embodiment, the magnetic core (M) comprises, preferably consists of, 1-20 magnetic nanoparticles and optionally a coating C2, i.e. magnetic nanoparticles, optionally with a coating C2, the nanoparticles forming said magnetic core (M) typically the magnetic core comprises 1-20 magnetic nanoparticles, preferably 1-10, more preferably 1-5, most preferably 1-3 nanoparticles.
Preferably, in this case, the nanoparticles comprise, more preferably consist of, a compound selected from the group consisting of metals, metal carbides, metal nitrides, metal sulfides, metal phosphides, metal oxides, metal chelates, and mixtures of two or more thereof it is to be understood that each nanoparticle may comprise, preferably consist of, a mixture of two or more of the foregoing groups, i.e., two or more of a metal, a metal carbide, a metal nitride, a metal sulfide, a metal phosphide, a metal oxide, a metal chelate, and a mixture of two or more thereof.
Thus, according to particularly preferred embodiments, the nanoparticles comprise a metal oxide, most preferably an iron oxide, in particular Fe3O4And more preferably consists thereof.
According to this embodiment, it is preferred that in case more than magnetic cores (M) are present in the magnetic particles, these magnetic cores (M) are not aggregated with each other.
Magnetic core comprising a nanoparticle (M)
According to a second preferred embodiment, the magnetic core (M) comprises more than 20 nanoparticles, and, typically, more than 100 nanoparticles, wherein these nanoparticles are preferably aggregated with each other to form a super-particle, more preferably, in this case, the magnetic core (M) comprises a super-particle consisting of aggregated, coated nanoparticles, preferably, in this case, the magnetic core (M) comprises a super-particle comprising 100-150 ten thousand nanoparticles, more preferably 750,000 nanoparticles, more preferably 1,750-320,000 nanoparticles, especially 90,000-320,000 nanoparticles, preferably, each nanoparticle is coated with at least coatings C2., preferably, in this case, the magnetic core (M) thus comprises, preferably consists of, a super-particle consisting of coated nanoparticles aggregated with each other, wherein the nanoparticles are coated with at least coatings C2, and wherein this coating is preferably deposited on the surface of the nanoparticles, preferably also with a super-coating C1.
Thus, according to this second preferred embodiment of the present invention, the magnetic particles according to the present invention comprise more than 20 magnetic nanoparticles, preferably 100-150 million nanoparticles, wherein said nanoparticles form at least super-particles each of which is typically coated with at least coating C2 and the super-particles are typically coated with at least coating C1.
Preferably, the coating C2 is a coating covering at least portions of each nanoparticle, preferably the entire surfaceIn preferred embodiments, the metal is a transition metal, preferred transition metals according to the invention include, but are not limited to, chromium, manganese, iron, cobalt, nickel, zinc, cadmium, nickel, gadolinium, copper, and molybdenum, most preferred the metal is iron, according to particularly preferred embodiments, each nanoparticle contained in a super-particle is a metal oxide nanoparticle, most preferred an iron oxide nanoparticle, particularly Fe3O4And (3) nanoparticles.
The present invention therefore also relates to magnetic particles as described above, wherein said magnetic core (M) comprises or preferably consists of a super-particle consisting of aggregated at least 20 magnetic nanoparticles, wherein said nanoparticles are preferably coated with at least coating layers C2, as well as to magnetic particles obtained or obtainable by the above-described method.
Preferably, the magnetic core (M) comprising the optional at least coating layers C1 has a diameter of 80-500 nm, more preferably 150-400nm, most preferably 200-300 nm, as determined according to DLS (ISO 22412).
Coating C2
As coating C2, generally any coating known to those skilled in the art may be considered, however, preferably coating C2 is selected from at least of the group consisting of dicarboxylic acids, tricarboxylic acids, polyacrylic acids, amino acids, surfactants, and fatty acids it is to be understood that the above group includes salts and derivatives of the compounds, such as esters and polymers, accordingly, coating C2 is preferably selected from at least of the group consisting of dicarboxylic acids, dicarboxylic acid salts, dicarboxylic acid derivatives, tricarboxylic acids, tricarboxylic acid salts, tricarboxylic acid derivatives, polyacrylic acids, polyacrylic acid salts, polyacrylic acid derivatives, amino acids, amino acid salts, amino acid derivatives, surfactants, salts of surfactants, fatty acids, fatty acid salts, and fatty acid derivatives.
As used herein, the term coated or coated is used to refer to the process of adsorption, van der waals forces, and/or nonpolar group interactions (e.g., chemisorption or physisorption) or covalent bonding between the magnetic nanoparticle or nanoparticle core and the coating C2 or C1 or two or more coatings, if present.
Preferably as a fatty acid, fatty acid salt or fatty acid derivative, a compound is selected which is capable of binding to the surface of the nanoparticles, thereby preferably stabilizing the nanoparticles. The fatty acid used as coating C2 is preferably a single-chain alkyl group having 8 to 22 carbon atoms, which has a terminal carboxyl group (-COOH) and a high affinity adsorption (e.g. chemisorption or physisorption) to the surface of the magnetic particles. Fatty acids serve multiple functions, including protecting the magnetic particle core from oxidation and/or hydrolysis in the presence of water, which can significantly reduce the magnetization of the nanoparticles (Hutten et al (2004)J. Biotech.112:47-63), stabilizing the nanoparticle core, and the like, the term "fatty acid" includes saturated or unsaturated fatty acids, particularly unsaturated fatty acids, exemplary saturated fatty acids include lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, propionic acid, butyric acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, decapetanoic acid, tridecylic acid, pentadecanoic acid, heptadecanoic acid, nonadecanoic acid, eicosanoic acid, docosanoic acid, tricosanoic acid, tetracosanoic acid, pentacosanoic acid, hexacosanoic acid, heptacosanoic acid, montanic acid, nonacosanoic acid, triacontanoic acid (hentriacontanoic acid), tridecanoic acid, tricosanoic acid, pentacosanoic acid (corosolic acid), triacontanoic acid, heptacosanoic acid, triacontanoic acid, and the likeFatty acids may be synthetic or may be isolated from synthetic fatty acids, i.e. fatty acid esters which may be substituted, i.e. fatty acid esters which may be isolated, i.e. fatty acid esters which may be substituted, i.e. fatty acid esters which may be obtained by reaction of synthetic or synthetic fatty acids, i.e. fatty acid esters which may be obtained by reaction of fatty acids, i.e. fatty acid esters which may be isolated, i.e. in particular in the form of fatty acid amides, , fatty acid esters, i.e. fatty acid esters which may be substituted, i.e. fatty acid esters which may be obtained by reaction of fatty acids such as fatty acid amides, e. fatty acid esters which may be obtained by reaction of fatty acids, e.g. fatty acid esters which may be obtained by reaction of synthetic or synthetic fatty acids, i.e. fatty acid esters, e. fatty acids which may be substituted, i.e. fatty acid esters which may be isolated from synthetic fatty acids, i.e. fatty acids such as fatty acid amides, fatty acid esters which may be substituted, e. fatty acid esters which may be obtained by reaction of synthetic or isolated, in particular in the case, fatty acids described herein, fatty acid esters, e. fatty acid esters, fatty acid esters which may be substituted, e. fatty acid esters which may be obtained by reaction of fatty acids, e. fatty acids (.
Surfactants used in the context of the present invention are amphiphilic, i.e. organic compounds containing hydrophobic and hydrophilic groups, preferably surfactants capable of binding to the surface of the ultragranulate, thereby preferably stabilizing the ultragranulate, non-limiting examples of surfactants having various chain lengths, hydrophilic-lipophilic balance (HLB) values and surface charges may be used, depending on the application, preferably the surfactants according to the present invention are quaternary ammonium salts, alkyl benzene sulfonates, lignosulfonates, polyoxyethoxylates (polyoxyethylalkoxylates) or sulfates, cetyl trimethylammonium bromide, cetyl trimethylammonium chloride, nonylphenol polyethoxylates (i.e. NP-4, NP-40 and NP-7), sodium dodecylbenzene sulfonate, ammonium lauryl sulfate, laureth sodium lauryl sulfate, myrith sodium sulfate, sodium docusate, perfluorooctanesulfonate, perfluorobutane sulfonates, alkyl-aryl ether phosphates, alkyl ether phosphates, sodium stearate, 2-acrylamido-2-methylpropane sulfonate, polyoxyethylene lauryl sulfate, polyoxyethylene lauryl ether (laureth) sulfate, polyoxyethylene lauryl betaine, polyoxyethylene sorbitan fatty alcohol, polyoxyethylene sorbitan fatty acid, polyoxyethylene ether, polyoxyethylene sorbitan fatty acid, polyoxyethylene ether, polyoxyethylene sorbitan fatty acid, polyoxyethylene ether, polyoxyethylene lauryl fatty acid, polyoxyethylene sorbitan fatty acid, polyoxyethylene ether, polyoxyethylene lauryl betaine, polyoxyethylene sorbitan fatty acid, polyoxyethylene lauryl fatty acid, polyoxyethylene ether, polyoxyethylene sorbitan fatty acid, polyoxyethylene lauryl betaine, polyoxyethylene lauryl fatty acid, polyoxyethylene ether, polyoxyethylene lauryl fatty acid, polyoxyethylene lauryl ether, polyoxyethylene lauryl betaine, polyoxyethylene ether, polyoxyethylene lauryl betaine, polyoxyethylene lauryl fatty acid, polyoxyethylene lauryl betaine, polyoxyethylene lauryl sulfate, polyoxyethylene lauryl betaine, polyoxyethylene lauryl ether, polyoxyethylene lauryl fatty acid, polyoxyethylene lauryl betaine, polyoxyethylene.
The term "amino acid" as used within the meaning of the present invention refers to natural or unnatural amino acids or amino acid derivatives as well as salts of amino acids. Preferably, the amino acids are selected to be capable of binding to the surface of the nanoparticles, thereby preferably stabilizing the nanoparticles. Exemplary amino acids include cysteine, methionine, histidine, alanine, arginine, asparagine, aspartic acid, glutamic acid, glutamine, glycine, isoleucine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, selenocysteine, pyrrolysine, cysteine, dehydroalanine, endosidinine, lanthionine, norvaline, and derivatives thereof.
The term "dicarboxylic acid" in the meaning of the present invention refers to a hydrocarbon or substituted hydrocarbon containing two carboxylic acid functional groups (i.e. R)1-(C(O)OH)2) Wherein R is1Representative dicarboxylic acids are, for example, malonic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, suberic acid, azelaic acid, sebacic acid, deca alkanedioic acid, dodecanedioic acid, hexadecanedioic acid, maleic acid, fumaric acid, glutaconic acid, callus acid, muconic acid, pentynedioic acid, citraconic acid, mesaconic acid, malic acid, aspartic acid, glutamic acid, tartronic acid, tartaric acid, diaminopimelic acid, glucaric acid, mesooxalic acid, oxaloacetic acid, acetonedicarboxylic acid, arabinic acid (arabinaric acid), phthalic acid, terephthalic acid, isophthalic acid, biphenyldicarboxylic acid, 2, 6-naphthalenedicarboxylic acid.
The term "tricarboxylic acid" in the sense of the present invention means a hydrocarbon or substituted hydrocarbon containing three carboxylic acid functions (i.e. R)1-(C(O)OH)3) Wherein R is1Is (a) a straight-chain hydrocarbon having 3 to 18 carbon units or (b) a cyclic hydrocarbon having 3 to 8 carbon units, which is an aromatic or non-aromatic ring. The term includes salts and derivatives of fatty acids, such as esters of fatty acids. Representative tricarboxylic acids are, for example, citric acid (2-hydroxypropane-1, 2, 3-tricarboxylic acid), isocitric acid (1-hydroxypropane-1, 2, 3-tricarboxylic acid), aconitic acid (prop-1-ene-1, 2, 3-trimethyltricarboxylic acid)Acids), propane-1, 2, 3-tricarboxylic acid, trimellitic acid (benzene-1, 2, 4-tricarboxylic acid), trimesic acid (benzene-1, 3, 5-tricarboxylic acid), oxalyl succinic acid (1-oxopropane-1, 2, 3-tricarboxylic acid) or hemimellitic acid (benzene-1, 2, 3-tricarboxylic acid). Preferably, the tricarboxylic acid is citric acid, including citrate salts, i.e. salts and derivatives of citric acid.
Preferably, C2 is selected from citric acid, histidine, CTAB, CTAC, sodium oleate, polyacrylic acid or a mixture of two or more thereof (including respective salts or derivatives thereof) thus, the invention also relates to magnetic particles as described above, and magnetic particles obtained or obtainable by the above method, wherein the magnetic core (M) preferably consists of super particles consisting of aggregated magnetic nanoparticles having at least coating C2, wherein the at least coating C2 is selected from citrate, histidine, CTAB, CTAC, sodium oleate, polyacrylic acid or a mixture of two or more thereof.
Preferably, the amount of coating C2 is 1 to 80 wt%, more preferably 5 to 70 wt%, more preferably 10 to 50 wt%, most preferably 20 to 40 wt%, based on the total weight of the sum of C2 and the nanoparticles.
Coating C1
The present invention thus also relates to magnetic particles as described above, and to magnetic particles obtained or obtainable by the above method, wherein at least magnetic cores (M) further comprise a coating C1.
The coating C1 is preferably deposited on the surface of the magnetic core (M) it will be appreciated that between the coating C1 and the magnetic core (M) there may be an additional separation layer, however, according to preferred embodiments, the C1 is coated directly on the magnetic core (M).
Preferably, the coating C1 surrounds the entire surface of the magnetic core (M).
In principle, any suitable coating known to the person skilled in the art may be used. Preferably, coating C1 is selected from the group consisting of surfactants, silica, silicates, silanes, phosphates, phosphonates, phosphonic acids, and mixtures of two or more thereof.
The present invention therefore also relates to a magnetic particle as described above, and to a magnetic particle obtained or obtainable by the above method, comprising at least magnetic cores (M), wherein the at least magnetic cores (M) comprise at least coating C1, and wherein the coating C1 is selected from the group consisting of surfactants, silica, silicates, silanes, phosphates, phosphonates, phosphonic acids and mixtures of two or more thereof.
Preferably, coating C1 is selected from the group consisting of silica, tetraethyl orthosilicate, 3- (trimethoxysilyl) propyl methacrylate, vinyltrimethoxysilane, vinyltriethoxysilane, allyltrimethoxysilane, allyltriethoxysilane, triethoxyvinylsilane, 3- (trimethoxysilyl) propyl acrylate, trimethoxy (7-octen-1-yl) silane, trimethoxymethylsilane, triethoxymethylsilane, ethyltrimethoxysilane, triethoxy (ethyl) silane, trimethoxyphenylsilane, trimethoxy (2-phenylethyl) silane, trimethoxy (propyl) silane, n-propyltriethoxysilane, isobutyl (trimethoxy) silane, isobutyltriethoxysilane, vinylphosphonic acid, dimethyl vinylphosphonate, diethyl allylphosphonate, diethyl allylphosphate, (2-methallyl) phosphonate, octylphosphonic acid, butylphosphonic acid, decylphosphonic acid, hexylphosphonic acid, hexadecylphosphonic acid, n-dodecylphosphonic acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, propionic acid, butyric acid, caproic acid, heptanoic acid, caprylic acid, capric acid, tridecanoic acid, eicosanoic acid, docosenoic acid, eicosanoic acid, docosenoic acid, eicosanoic acid, docosenoic acid, eicosapentaenoic acid, eicosapentad, eicosapentaenoic acid, eicosapentad, eicosapentanoic acid, eicosapentad.
Preferably, each magnetic core (M) comprises a coating C1 in an amount of 1 to 40 wt. -%, preferably 2 to 15 wt. -%, more preferably 5 to 10 wt. -%, based on the total weight of at least magnetic cores (M).
According to preferred embodiments of the present invention, coating C1 comprises vinyl or acryl groups.
Polymer matrix (P)
As mentioned above, each particle comprises a polymer matrix (P) in addition to at least magnetic cores (M).
Preferably, the polymer matrix (P) is a porous polymer matrix, preferably comprising a porous polymer matrix having pores with a pore diameter of less than 100 nm, more preferably less than 90 nm, more preferably less than 80 nm, more preferably less than 70 nm, more preferably less than 60 nm, more preferably less than or equal to 50 nm, e.g. 0.5 nm to 50 nm, preferably 1 to 20 nm, as determined according to ISO 15901.
The present invention therefore also relates to a magnetic particle as described above, and to a magnetic particle obtained or obtainable by the above-described process, wherein the polymer matrix (P) is a porous polymer matrix comprising pores having a pore size of less than 100 nm, preferably less than or equal to 50 nm, as determined according to ISO 15901.
Preferably, at least 90% of all pores present in the polymer matrix have a pore size of less than 10 nm and at least 50% of all pores present in the polymer matrix have a pore size of less than 5 nm, as determined according to ISO 15901.
According to particularly preferred embodiments, the polymer matrix does not comprise macropores, i.e. pores having a pore diameter of more than 50 nm.
Preferably, the particles comprise the polymer matrix (P) in an amount of from 40 to 98 wt. -%, more preferably from 50 to 95 wt. -%, more preferably from 60 to 90 wt. -%, most preferably from 70 to 85 wt. -%, based on the total weight of the particles.
The polymer matrix (P) preferably comprises a copolymer obtained or obtainable by a process comprising the polymerisation of at least two different monomer building blocks selected from styrene, functionalised styrene, vinylbenzyl chloride, divinylbenzene, vinyl acetate, methyl methacrylate and acrylic acid. Preferably, the copolymer obtained or obtainable by a process comprising the polymerisation of at least two different monomer building blocks selected from the following monomers:
wherein R is1、R2、R3、R4And R5Independently of one another from the group-N3、-NH2-Br, -I, -F, -NR ' R ' ', -NR ' R ' ' ', -COOH, -CN, -OH, -OR ', -COOR ', -NO2, -SH2, -SO2, -R ' (OH) x, -R ' (COOH) x, -R ' (COOR ' ') x, -R ' (OR ' ') x, -R ' (NH2) x, -R ' (NHR ' ') x, -R ' (NR "R ' ' ') x, -R ' (Cl) x, -R ' (I) x, -R ' (Br) x, -R ' (F) x, R ' (CN) x, -R ' (N3) x, -R ' (NO2) x, -R ' (SH2) x, -R ' (SO2) x, alkyl, aryl, cycloalkyl, heteroaryl, N ' (N) x, Heterocycloalkyl, wherein R ', R "and R' '' are independently from each other selected from alkyl, aryl, cycloalkyl, heteroaryl, heterocycloalkyl, halide, hydrogen, thioether, nitrate and amine, and wherein x is an integer from 1 to 3.
Preferably, the polymer matrix comprises a crosslinked polymer, more preferably obtained or obtainable by a process comprising copolymerizing suitable monomeric building blocks, which are crosslinking agents, and thus agents for effecting crosslinking therewith in the resulting polymer, in the presence of at least monomeric building blocks suitable agents for crosslinking the polymer are known to those skilled in the art and include, but are not limited to, building blocks such as divinylbenzene, bis (vinylphenyl) ethane, bis (vinylbenzyloxy) hexane, bis (vinylbenzyloxy) dodecane, and derivatives thereof.
According to particularly preferred embodiments, divinylbenzene is used as crosslinking agent.
Preferably, the polymer matrix is obtained or obtainable by a process comprising copolymerizing monomer building blocks, wherein 5 to 90 volume% of all monomer building blocks are cross-linking agents.
Preferably, a degree of crosslinking of at least 5% is obtained in the resulting polymer.
The at least magnetic cores (M) are preferably embedded in the polymer matrix the term "embedded" in this context is meant to mean that the magnetic cores are preferably completely surrounded by the polymer matrix or that they may be partially surrounded by the polymer matrix.
As mentioned above, according to preferred embodiments, the particles comprise at least two magnetic cores (M). in this case, it will be understood that each magnetic core (M) present in the particle is embedded in the polymer matrix (P.) thus, the invention also relates to a magnetic particle as described above, as well as to a magnetic particle obtained or obtainable by the above method, wherein the at least two magnetic cores (M) are embedded in the polymer matrix.
Hypercrosslinking
More preferably, the polymer matrix P comprises a crosslinked copolymer obtained or obtainable by a process comprising block polymerizing at least two different monomer building blocks as described above, thereby preferably obtaining a crosslinked polymer, wherein said crosslinked polymer is further hypercrosslinked at step .
The term "hypercrosslinking" as used herein refers to multiple crosslinks of the type produced in a rigid three-dimensional network preferably, hypercrosslinking is achieved by subjecting a crosslinked polymer to a chemical reaction, thereby obtaining a hypercrosslinked polymer, thus, the polymer matrix (P) is a polymer matrix obtained or obtainable by hypercrosslinking the polymer steps by a chemical reaction, with or without a hypercrosslinking agent suitable agents for hypercrosslinking the polymer are known to those skilled in the art and include, but are not limited to, dichloroethane, dichloromethane, chloroform, carbon tetrachloride, dichlorobenzene, trichlorobenzene, dichloroalkane, dibromoalkane, diiodoalkane, trichloroalkane, tribromoalkane, triiodoalkane, dimethoxymethane, dimethoxyethane and mixtures of two or more thereof.
Preferably, hypercrosslinking is achieved by subjecting the crosslinked polymer matrix to a friedel-crafts reaction, in particular a friedel-crafts reaction as described herein below.
Surface functionalization
It is understood that a magnetic particle comprising at least magnetic cores M and a polymer matrix (P) may further comprise steps of surface modification of the surface of the particle, and thus the surface of the polymer matrix (P) is preferably functionalized with at least groups selected from-OH, -COOH, diethylaminoethanol, R-SO2-OH、-NH2、R-SO2-OH、-RNH、-R2N、-R3N+-CH3、-C2H5、-C4H9、-C8H17、-C18H37、-C6H5、-C6H9NO6Phenyl-hexyl, biphenyl, hydroxyapatite, boronic acid, biotin, azide, epoxide, alkyl, aryl, cycloalkyl, heteroaryl, heterocycloalkyl, amino acid, -COOR, -COR, -OR, antibodies and fragments thereof, aptamers, nucleic acids, and receptor proteins OR binding domains thereof. Preferably, these groups are covalently attached to suitable functions of the polymer matrixCan be clustered. Methods for performing such modifications are known to those skilled in the art.
Method for producing magnetic particles
As mentioned above, the present invention also relates to a method for the preparation of a magnetic particle comprising a polymer matrix (P) and at least magnetic cores (M), preferably at least two magnetic cores (M), wherein the polymer matrix (P) comprises at least cross-linked polymers, wherein the magnetic particle has a particle size in the range of 1-60 micrometer, the method comprising:
(i) providing at least magnetic cores (M), preferably at least two magnetic cores (M),
(ii) providing a precursor molecule of the polymer,
(iii) (iii) polymerizing the polymer precursor molecules according to (ii) in the presence of at least magnetic cores (M) thereby forming particles comprising at least magnetic cores (M), preferably at least two magnetic cores (M), which are embedded in a polymer matrix (P1), wherein the polymer matrix (P1) preferably comprises, more preferably consists of, a cross-linked polymer, and
(iv) (iv) preferably hypercrosslinking the polymer matrix (P1) of the particles obtained in (iii),
to obtain magnetic particles.
Preferably in (iii), at least magnetic cores (M) are embedded in the matrix.
Step (i)
As mentioned above, the at least magnetic cores (M) preferably comprise a compound selected from the group consisting of a metal, a metal carbide, a metal nitride, a metal sulfide, a metal phosphide, a metal oxide, a metal chelate and a mixture of two or more thereof the at least magnetic cores (M) may further comprise an alloy with a metal such as gold, silver, platinum or copper more preferably the at least magnetic cores (M) comprise a metal oxide or a metal carbide, more preferably the at least magnetic cores (M) comprise an iron oxide, in particular an iron oxide selected from the group consisting of Fe3O4、α-Fe2O3、γ- Fe2O3、MnFexOy、CoFexOy、NiFexOy、CuFexOy、ZnFexOy,、CdFexOy、BaFexO and SrFexO, wherein x and y vary depending on the synthesis method, and wherein x is preferably an integer from 1 to 3, more preferably 2, and wherein y is preferably 3 or 4, most preferably, the at least magnetic cores (M) comprise Fe3O4。
Thus, the present invention also relates to a method as described above, and to magnetic particles obtained or obtainable by said method, wherein said at least magnetic cores (M) comprise a metal oxide or metal carbide, more preferably said at least magnetic cores (M) comprise an iron oxide, in particular an iron oxide selected from the group consisting of Fe3O4、α-Fe2O3、γ-Fe2O3、MnFexOy、CoFexOy、NiFexOy、CuFexOy、ZnFexOy,、CdFexOy、BaFexO and SrFexO, wherein x and y vary depending on the synthesis method, and wherein x is preferably an integer from 1 to 3, more preferably 2, and wherein y is preferably 3 or 4, most preferably, the at least magnetic cores (M) comprise Fe3O4。
As mentioned above, the magnetic core (M) preferably comprises, more preferably consists of, magnetic nanoparticles and a coating C1.
Preferably, step (i) comprises:
(i.1) providing at least magnetic nanoparticles, and
(i.2) coating the at least magnetic nanoparticles with a coating C1, the coating C1 preferably being selected from the group consisting of surfactants, silica, silicates, silanes, phosphates, phosphonates, phosphonic acids and mixtures of two or more thereof
To obtain a magnetic core (M).
The magnetic nanoparticles in (i.1) comprise, preferably consist of, at least magnetic nanoparticles, preferably superparamagnetic nanoparticles, and optionally a coating C2.
Thus, step (i.1) comprises providing at least nanoparticles.
Methods for providing magnetic nanoparticles are known to those skilled in the art and are described, for example, in Lu, Salabas, Sch ü th, angelw Chem int. ed. 2007, 46,1222-1244, the contents of each of which are incorporated herein by reference.
Especially in Fe3O4In the case of nanoparticles, fe (ii) and fe (iii), the oxides precipitate from an aqueous medium, preferably a basic aqueous medium, to give the respective nanoparticles.
According to the above-mentioned th preferred embodiment, wherein each magnetic core (M) comprises at least magnetic nanoparticles, optionally with a coating C1, step (i.1) thus comprises providing at least magnetic nanoparticles, optionally with a coating C1, forming the magnetic core (M), preferably, in this case the magnetic nanoparticles do not comprise a coating C2. according to this preferred embodiment of the invention, the magnetic particles according to the invention preferably comprising 1-20 magnetic nanoparticles.
According to this embodiment, it is preferred that in case more than magnetic cores (M) are present in the magnetic particles, these magnetic cores (M) are not aggregated with each other.
Accordingly, the present invention also relates to a method as described above, and to magnetic particles as described above, wherein step (i) comprises:
(i.1) providing at least magnetic nanoparticles optionally having a coating C2,
obtaining at least magnetic nanoparticles, an
(i.2) coating the at least magnetic nanoparticles with a coating C1, coating C1 preferably being selected from the group consisting of surfactants, silica, silicates, silanes, phosphates, phosphonates, phosphonic acids and mixtures of two or more thereof
To obtain a magnetic core (M).
According to the above second preferred embodiment the magnetic core (M), i.e. preferably each magnetic core (M), comprises more than 20 nanoparticles, preferably 100-150 ten thousand nanoparticles, more preferably 750-750,000 nanoparticles, more preferably 1,750-320,000 nanoparticles, especially 90,000-320,000 nanoparticles, more preferably the nanoparticles form at least super-particles consisting of aggregated, coated nanoparticles preferably the super-particles are coated with at least coatings C1 preferably in this case the super-particles are coated with at least coatings C1, wherein the coatings are preferably deposited on the surface of the super-particles, hence the super-particles are preferably coated with a coating C1 it is to be understood that each of the super-particles is preferably coated with e.g. a coating C2. step (i.1) thus comprising providing more than 20 nanoparticles, wherein the method further comprises aggregating the nanoparticles to form at least 2 super-particles, and optionally at least 387C super-particles C64 per coating 1.
Accordingly, the present invention also relates to a method as described above, and to magnetic particles as described above, wherein step (i) comprises:
(i.1) providing at least superparticles by
(i.1.1) providing more than 20 nanoparticles, preferably with a coating C2,
(i.1.2) aggregating the nanoparticles, thereby forming a nanoparticle, and
(i.2) coating the at least nanoparticles with a coating C1, the coating C1 preferably being selected from the group consisting of surfactants, silica, silicates, silanes, phosphates, phosphonates, phosphonic acids and mixtures of two or more thereof
To obtain a magnetic core (M).
In this case, each magnetic core (M) comprises a super-particle, which super-particle preferably consists of at least 21 coated nanoparticles, preferably 100-150 ten thousand nanoparticles, more preferably 750-750,000 nanoparticles, more preferably 1,750-320,000 nanoparticles, especially 90,000-320,000 nanoparticles, which are coated and aggregated with each other.
It will be appreciated that (i.1.1) and (i.1.2) may be carried out in a single step by a reaction in which the nanoparticles are provided directly therein.
Methods of providing nanoparticles are known to those skilled in the art and are described, for example, in Liu et al, angelw.chem. int. ed. 2009, 48, 5875-.
Especially in the presence of Fe3O4In the case of nanoparticles of superparticle, the synthesis may include partial reduction of FeCl3Fe is preferably obtained at elevated temperature, e.g. at a temperature of 150 to 250 ℃ and preferably at elevated pressure, e.g. a pressure of 2-10 bar3O4-a nanoparticle.
As mentioned above, the coating C2 is thus selected from at least of dicarboxylic acids, dicarboxylic acid salts, dicarboxylic acid derivatives, polyacrylic acid salts, polyacrylic acid derivatives, tricarboxylic acids, tricarboxylic acid salts, tricarboxylic acid derivatives, amino acids, amino acid salts, amino acid derivatives, surfactants, salts of surfactants, fatty acids, fatty acid salts and fatty acid derivatives.
Methods of coating nanoparticles or nanoparticles are known to those skilled in the art. During the synthesis of the nanoparticles, the coating is preferably carried out in situ.
Step (ii)
In step (ii), polymer precursor molecules, i.e. monomer building blocks, are provided, which after polymerization result in the respective polymers.
Preferably, the polymer precursor molecules in (ii) are selected from styrene, functionalized styrene, vinylbenzylchloride, divinylbenzene, vinyl acetate, methyl methacrylate and acrylic acid, more preferably from the following monomers:
wherein R is1、R2、R3、R4And R5Independently of one another from the group-N3、-NH2、-Br、-I、-F、-NR'R"、-NR'R"R'''、-COOH、-CN、-OH、-OR'、-COOR'、-NO2、-SH2、-SO2、-R'(OH)x、-R'(COOH)x、-R'(COOR")x、-R'(OR")x、-R'(NH2)x、-R'(NHR")x、-R'(NR"R''')x、-R'(Cl)x、-R'(I)x、-R'(Br)x、-R'(F)x、R'(CN)x、-R'(N3)x、-R'(NO2)x、-R'(SH2)x、-R'(SO2)xAlkyl, aryl, cycloalkyl, heteroaryl, heterocycloalkyl, wherein R ', R "and R'" are independently from each other selected from alkyl, aryl, cycloalkyl, heteroaryl, heterocycloalkyl, halide, hydrogen, thioether, nitrate, and amine.
Step (iii)
In step (iii) the polymer precursor molecules according to (ii) are polymerized in the presence of at least magnetic cores (M) thereby forming particles comprising at least magnetic cores (M), preferably at least two magnetic cores (M), which are embedded in a polymer matrix (P1), wherein the polymer matrix (P1) preferably comprises, more preferably consists of, a cross-linked polymer as described above and below, then preferably the cross-linked polymer matrix P1 is further hyper-crosslinked in step (iv) resulting in polymer matrix P.
(iii) The term "suspension polymerization" refers to systems in which relatively water-insoluble polymerization precursor molecules are suspended as droplets in an aqueous phase, typically, a suspending agent is used to maintain suspension, and the resulting polymer is obtained as a dispersed solid phase, although monomer building blocks can be directly dispersed in suspension polymerization systems, hydrocarbon solvents or diluents are typically used with the monomer , such as n-heptane, isooctane, cyclohexane, benzene, toluene, and the like, including mixtures.
In a suspension polymerization system, the monomer mixture to be polymerized generally comprises the monomers, or, if desired, a monomer-in-monomer solution, the at least magnetic cores (M), solvent, and, if used, initiator.
Thus, step (iii) preferably comprises:
(iii.1) providing a composition (A) comprising a polymer precursor molecule according to (ii), at least magnetic cores (M) according to (i), at least organic solvents, at least initiators and an aqueous phase, wherein the organic solvents are immiscible with water, and optional additives, and
(iii.2) stirring the composition (A) to obtain an emulsion (B), wherein the emulsion is preferably an organic solvent-in-water emulsion.
Preferably, the polymerization in (iii) is carried out in the presence of an initiator selected from the group consisting of: azobis (isobutyronitrile) (AIBN), 2 '-azobis (2-methylbutyronitrile) (VAZO 67), 1' -azobis (cyanocyclohexane) (VAZO 88), Benzoyl Peroxide (BPO), 2 '-azobis (2-amidinopropane) dihydrochloride (AAPH) and 4, 4' -azobis (4-cyanopentanoic acid) (ACVA).
The monomer and at least magnetic cores (M) are preferably suspended in an aqueous solution optionally containing at least suspending agents the amount of water used can vary widely depending on the type of reactor used, the stirring means, etc., although the final suspension mixture preferably contains about 5-60 wt% monomer building blocks based on the total weight of the entire mixture including water.
In suspension polymerization systems, various suspending agents may be used as additives because the process involves a dispersion of liquid-in-liquid and provides the final product in the form of discrete solid particles. Suspending agents include insoluble carbonates, silicates, talc, gelatin, pectin, starch, cellulose derivatives, insoluble phosphates, PVA, salts, NaCl, KCl, PVP and the like. Preferably, the polymerization in (iii) is carried out in the absence of any surfactant.
The time taken for the polymerization should be sufficient to achieve the desired degree or degree of conversion and can vary within wide limits, depending on the various reaction parameters, such as the temperature used, from a few minutes to several hours, for example 48 hours. Preferably, step (iii) is carried out for a period of from 1 hour to 30 hours, preferably from 1 hour to 8 hours.
The temperature used is at least sufficient to complete the thermal polymerization, or to cause decomposition of the free radical initiator (if used), which provides initiation of the reaction, preferably below a temperature that may cause formation of a polymer gel. Preferably, temperatures of about 0 ℃ to 100 ℃, preferably 40 to 90 ℃ are used.
The stirring is preferably carried out with an overhead stirrer.
Preferably, the polymer matrix (P1) comprises a cross-linked polymer, more preferably obtained or obtainable by copolymerizing the polymer with a cross-linking agent. Suitable agents for crosslinking the polymer are known to those skilled in the art and include, but are not limited to, divinylbenzene, bis (vinylphenyl) ethane, bis (vinylbenzyloxy) hexane, bis (vinylbenzyloxy) dodecane, and derivatives thereof.
Preferably, the polymer matrix (P1) is obtained or obtainable by crosslinking the polymer with 5 to 90% by volume of a crosslinking agent, based on the total amount of polymer.
Step (iv)
The polymer matrix (P1) is preferably hypercrosslinked in a further step (iv) if no further step (iv) is performed, said polymer matrix (P1) preferably corresponds to a polymer, however, according to specific preferred embodiments, the hypercrosslinking in step (iv) is preferably performed by a Friedel-crafts reaction for the purposes of this application, the term "Friedel-crafts reaction" refers to the well known type of reaction developed by Charles Friedel and James crafts to attach substituents to aromatic rings by electrophilic aromatic substitution and includes the two main types of Friedel-crafts reaction: alkylation and acylation.
Preferably, (iv) is carried out in the presence of a catalyst comprising a lewis acid selected from the group consisting of: FeCl3、ZnCl2、AlCl3、BF3、SbCl5、SnCl4、TiCl4、SiCl4And mixtures of two or more thereof, or consist thereof. More preferably, the catalyst comprises FeCl3Or ZnCl2Or a mixture thereof, more preferably consisting thereof.
Preferably, the hypercrosslinking in (iv) is carried out at a temperature in the range of-30 ℃ to 120 ℃, more preferably the hypercrosslinking in (iv) is carried out at a temperature less than or equal to 80 ℃, such as in the range of-30 ℃ to 80 ℃.
More preferably (iv) is carried out at a temperature of less than 70 ℃, more preferably less than 60 ℃, more preferably less than 50 ℃, more preferably less than 40 ℃, more preferably less than 30 ℃, for example preferably at a temperature of from-30 ℃ to 30 ℃, more preferably at a temperature of from-20 ℃ to 30 ℃, more preferably at a temperature of from-10 ℃ to 30 ℃, more preferably at a temperature of from 0 ℃ to 30 ℃, most preferably at a temperature of from 10 ℃ to 30 ℃. The temperature may be varied constantly or stepwise, or kept substantially constant during the reaction. In the case of temperature variations, it is understood that the temperature is always maintained at a temperature equal to or less than 120 ℃, less than 100 ℃, less than 80 ℃, and preferably at a temperature less than 70 ℃, more preferably less than 60 ℃, more preferably less than 50 ℃, more preferably less than 40 ℃, more preferably less than 30 ℃, for example preferably at a temperature of-30 ℃ to 30 ℃, more preferably at a temperature of-20 ℃ to 30 ℃, more preferably at a temperature of-10 ℃ to 30 ℃, more preferably at a temperature of 0 ℃ to 30 ℃, most preferably at a temperature of 10 ℃ to 30 ℃.
The crosslinking may be carried out by any suitable solvent known to the skilled person, preferably in a solvent comprising dichloroethane, toluene, acetonitrile, DMF, diethyl ether, THF, benzene, xylene, dioxane, an alkane, dichloromethane, chloroform, chlorobenzene, carbon tetrachloride, NMP, dichlorobenzene, trichlorobenzene, an ether, a cycloalkane, an organic halide or a mixture of two or more thereof. More preferably, the crosslinking in (iv) is carried out in a solvent selected from dichloroethane, toluene, acetonitrile, DMF, diethyl ether, THF, benzene, xylene, dioxane, alkane, dichloromethane, chloroform, chlorobenzene, carbon tetrachloride.
Preferably, the reaction in (iv) is not carried out in dichloroethane or other organic halide, more preferably the reaction in (iv) is not carried out in a solvent comprising dichloroethane or other organic halide.
In particular, the reaction in (iv) is carried out in a solvent comprising at least THF, acetonitrile, DMF, dioxane or toluene. More preferably, (iv) is carried out in a solvent selected from THF, acetonitrile, DMF, dioxane, toluene and mixtures of two or more thereof.
Preferably, the reaction in (iv) is carried out for a reaction time of 4 hours or less, for example, 10 minutes to 2 hours, more preferably 30 minutes to 1.5 hours, more preferably 45 minutes to 1 hour.
Preferably, the reaction in (iv) is carried out under an inert atmosphere, even more preferably during the reaction in (iv), an inert gas is flowed through the mixture. Preferably, the inert gas is nitrogen and/or argon.
Optional step (v)
In addition to the above steps, the method may comprise or more additional steps in particular, the method may comprise functionalizing the surface of the polymer particles obtained according to (iii) or (iv).
Accordingly, the present invention also relates to a method as described above, and to magnetic particles obtained or obtainable by the above method, wherein the method further comprises the step of:
(i) (iv) functionalizing the surface of the polymer particles according to (iii) or (iv).
Preferably in this step, the polymer particles are functionalized with a group selected from: -OH, -COOH, diethylaminoethanol, R-SO2-OH、-NH2、R-SO2-OH、-RNH、-R2N、-R3N+-CH3、-C2H5、-C4H9、-C8H17、-C18H37、-C6H5、-C6H9NO6Phenyl-hexyl, biphenyl, hydroxyapatite, boronic acid, biotin, azide, epoxide, alkyl, aryl, cycloalkyl, heteroaryl, heterocycloalkyl, amino acid, -COOR, -COR, -OR, antibodies and fragments thereof, aptamers, nucleic acids, polymers, and receptor proteins OR binding domains thereof.
Methods for functionalizing polymeric particles are known to those skilled in the art and are described, for example, in Bioconjugate technologies, second edition, g.t. hermanson, the disclosure of which is incorporated herein by reference.
Preferably, this step comprises at least treating the particles with a suitable base (OH functionalization), such as KOH.
Use/method of analysis
The above-described magnetic particles and the magnetic particles obtainable or obtainable by the above-described method are preferably used for the qualitative and/or quantitative determination of at least analytes in a fluid.
The term "qualitative" determination as used herein refers to the determination of the presence or absence of at least analytes in a fluid furthermore, the term may also include the evaluation of the nature of the analytes, i.e. it may include the identification of the analytes or the identification of the class of chemical molecules to which the analytes belong.
To determine whether an analyte binds to a magnetic particle, the compound bound to the particle may be eluted by a suitable technique and the presence or absence of the at least analyte in the eluate may then be determined.
The at least analytes or chemical classes to which they belong can be identified by suitable analytical methods, such as mass spectrometry, UV-vis, NMR, IR or biochemical methods such as ELISA, RIA etc. after the analytes have been eluted from the magnetic particles.
The term "quantifying" as used herein refers to determining the absolute or relative amount of at least analytes contained in a fluid sample.
The amount of at least analytes can be determined as described above for a qualitative determination, however, after the analytes are eluted from the magnetic particles, it is desirable to determine the amount in the eluate.
In view of the above, the present invention also contemplates a method for determining at least analytes in a fluid sample, comprising the steps of:
(a) contacting the magnetic particles according to the invention or the magnetic particles obtained or obtainable by the method of the invention with a fluid sample comprising or suspected to comprise said at least analytes, and
(b) determining at least analytes eluted from the magnetic particles.
Typically, the assay referred to in this context is a qualitative or quantitative assay.
Typically, step (a) of the method is performed for a time and under conditions sufficient to allow at least analytes to bind to the magnetic particles.
Preferably, step (a) further comprises the steps of:
(a1) preferably without eluting the at least analytes, to which at least moieties of at least analytes are bound, and/or
(a2) Eluting the at least analytes from the magnetic particles under conditions suitable to allow elution of the at least analytes.
More specifically, the qualitative or quantitative determination in (b) may comprise determining the presence or absence of bound analyte on the magnetic particles, or determining the amount of analyte bound to the magnetic particles.
It will be appreciated that the washing step in (a1) may be performed as a single washing step or, alternatively, more than washing steps may be performed.
More specifically, the qualitative determination may include the following additional steps as part of steps (a) and/or (b):
-determining whether the at least analytes are bound to the magnetic particles.
The use of the magnetic particles of the invention or obtainable by the method of the invention advantageously reduces matrix carryover of fluids in applications as described above.
The analyte determined by the magnetic particles of the invention or by the magnetic particles obtained by the method of the invention or the analyte determined according to the above-mentioned use is preferably a chemical compound present in a solution of a biological fluid sample, an environmental sample or a mixture of chemical compounds. Thus, the fluid sample according to the present invention is preferably selected from the group of fluids consisting of: a body fluid, a liquid or dissolved environmental sample, and a solution of a mixture of chemical compounds.
In preferred embodiments, a fluid sample as used herein refers to a biological sample obtained for the purpose of in vitro evaluation.
Preferred fluid samples are whole blood, serum, plasma, bronchoalveolar lavage (BAL), Epithelial Lining Fluid (ELF), urine or sputum, with plasma or serum being most preferred.
The term fluid sample includes a biological sample that has been treated in any way after it has been obtained, for example by treating, solubilizing or enriching certain components, such as proteins or polynucleotides, with a reagent. Typically, the fluid sample is a liquid sample.
The fluid sample may be, for example, whole blood, serum, antibodies recovered from a patient, or plasma the fluid sample is preferably whole blood, serum, or plasma in embodiments the sample is a clinical sample in another embodiments the sample is for diagnostic analysis.
Depending on the nature of the fluid sample, different classes of chemical compounds are to be detected. Preferably, the analyte according to the present invention may be selected from steroids, sugars, vitamins, drugs (drugs) including drugs (mediators) and drugs of abuse (drugs for abuse), organic compounds, proteins, nucleic acids and mixtures of two or more thereof. These analytes, of crucial importance, are present in biological samples. However, they may also be present in environmental samples.
The above-described applications for determining analytes in fluid samples may preferably be applied to or relate to diagnostic purposes, drug abuse testing, environmental control, food safety, quality control, purification, or manufacturing processes.
Preferably, the analyte is determined by mass spectrometry, UV-vis, NMR, IR.
Summarizing the findings of the present invention, the following embodiments are particularly preferred:
1. magnetic particle comprising a polymer matrix (P) and at least magnetic cores (M), preferably at least two magnetic cores, wherein the polymer matrix comprises at least cross-linked polymers, and wherein the magnetic particle has a particle size in the range of 1-60 microns as determined according to ISO 13320.
2. The magnetic particle of embodiment 1, wherein the polymer matrix comprises pores having a pore size of less than 100 nm, preferably less than or equal to 50 nm, as determined according to ISO 15901-3.
3. The magnetic particle of
4. The magnetic particle of
5. The magnetic particle of any of embodiments 1 through 4, wherein the magnetic particle has at least 1A m2/kg, preferably at least 10A m2Saturation magnetization per kg, as determined according to ASTM A894/A894M.
6. The magnetic particle of any of embodiments 1 through 5 wherein the at least magnetic cores (M) comprise a compound selected from the group consisting of a metal, a metal carbide, a metal nitride, a metal sulfide, a metal phosphide, a metal oxide, a metal chelate and a mixture of two or more thereof.
7. The magnetic particle of any of embodiments 1 through 6, wherein the at least magnetic cores (M) comprise a metal oxide or metal carbide, more preferably an iron oxide, in particular an iron oxide selected from the group consisting of Fe3O4、α-Fe2O3、γ- Fe2O3、MnFexOy、CoFexOy、NiFexOy、CuFexOy、ZnFexOy,、CdFexOy、BaFexO and SrFexO, wherein x and y vary depending on the synthesis method, and wherein x is preferably an integer of 1 to 3, more preferably 2, and wherein y is preferably 3 or 4, most preferably Fe3O4。
8. The magnetic particle of any of embodiments 1-7, wherein the magnetic particle is superparamagnetic.
9. The magnetic particle of any of embodiments 1-8, wherein the at least magnetic cores (M) comprise at least magnetic nanoparticles, preferably at least iron oxide nanoparticles, more preferably Fe3O4-nanoparticles.
10. The magnetic particle of embodiment 9, wherein the at least magnetic cores (M) comprise, more preferably consist of, magnetic nanoparticles and a coating C1.
11. The magnetic particle of
12. The magnetic particle of any of embodiments 1-8, wherein the at least magnetic cores (M) comprise, preferably consist of, nanoparticles and optionally comprise a coating C1.
13. The magnetic particle of
14. The magnetic particle of embodiment 13, wherein each nanoparticle is coated with at least coating C2, said coating preferably being deposited on the surface of the nanoparticle,
and wherein the coating C2 is preferably selected from the group consisting of dicarboxylic acids, dicarboxylic acid salts, dicarboxylic acid derivatives, tricarboxylic acids, tricarboxylic acid salts, tricarboxylic acid derivatives, amino acids, amino acid salts, amino acid derivatives, surfactants, salts of surfactants, polyacrylic acid salts, polyacrylic acid derivatives, fatty acids, fatty acid salts and fatty acid derivatives.
15. The magnetic particle of any of
16. The magnetic particle of any of
17. Magnetic particles of any of
18. The magnetic particle of
19. The magnetic particle of any of
20. The magnetic particle of any of
21. The magnetic particle of any of
22. The magnetic particle of any of embodiments 1 through 21 wherein the particle comprises at least two magnetic cores (M).
23. The magnetic particle of any of embodiments 1-22, wherein the polymer matrix (P) comprises a cross-linked copolymer obtained or obtainable by a process comprising copolymerizing suitable monomeric building blocks, which are cross-linking agents, in the presence of at least monomeric building blocks, wherein preferably 5-90vol% of all monomeric building blocks are cross-linking agents, more preferably divinylbenzene.
24. The magnetic particle of embodiment 23, wherein in the copolymerizing, a polymer having a degree of crosslinking of at least 5% is obtained.
25. The magnetic particle of any of embodiments 1-24, wherein the polymer matrix comprises a copolymer obtained or obtainable by a process comprising polymerization of at least two different monomer building blocks selected from the group consisting of styrene, functionalized styrene, vinylbenzylchloride, divinylbenzene, vinyl acetate, methyl methacrylate, and acrylic acid, wherein the monomer building blocks are preferably selected from the group consisting of:
wherein R is1、R2、R3、R4And R5Independently of one another from the group-N3、-NH2、-Br、-I、-F、-NR’R’’、-NR’R’’R’’’、-COOH、-CN、-OH、-OR’、-COOR’、-NO2、-SH2、-SO2、-R’(OH)x、-R’(COOH)x、-R’(COOR’’)x、-R’(OR’’)x、-R’(NH2)x、-R’(NHR’’)x、-R’(NR’’R’’’)x、-R’(Cl)x、-R’(I)x、-R’(Br)x、-R’(F)x、R’(CN)x、-R’(N3)x、-R’(NO2)x、-R’(SH2)x、-R’(SO2)xAlkyl, aryl, cycloalkyl, heteroaryl, heterocycloalkyl, and wherein R ', R ", and R'" are independently selected from alkyl, aryl, cycloalkyl, heteroaryl, heterocycloalkyl, halide, hydrogen, thioether, nitrate, and amine.
26. The magnetic particle of
27. The magnetic particle of any of embodiments 1 through 26 wherein at least magnetic cores (M), preferably at least two magnetic cores (M), are embedded in the polymer matrix.
28. The magnetic particle of any of embodiments 1 through 27 wherein the particle comprises at least two magnetic cores (M) and a polymer matrix (P).
29. The magnetic particle of any of embodiments 1 through 28 wherein the surface of the particle is functionalized with at least groups selected from the group consisting of-OH, -COOH, diethylaminoethanol, R-SO2-OH、-NH2、R-SO2-OH、-RNH、-R2N、-R3N+-CH3、-C2H5、-C4H9、-C8H17、-C18H37、-C6H5、-C6H9NO6Phenyl-hexyl, biphenyl, hydroxyapatite, boronic acid, biotin, azide, epoxide, alkyl, aryl, cycloalkyl, heteroaryl, heterocycloalkyl, amino acid, -COOR, -COR, -OR, antibodies and fragments thereof, aptamers, nucleic acids, and receptor proteins OR binding domains thereof.
30. The magnetic particle of any of embodiments 1 through 29, wherein the particle is a substantially spherical particle.
31. A method for the preparation of a magnetic particle comprising a polymer matrix (P) and at least magnetic cores (M), preferably at least two magnetic cores (M), wherein the polymer matrix (P) comprises at least crosslinked polymers, wherein the magnetic particle has a particle size in the range of 1-60 microns as determined according to ISO13320, the method comprising:
(i) providing at least magnetic cores (M), preferably at least two magnetic cores (M),
(ii) providing a precursor molecule of the polymer,
(iii) (iii) polymerizing the polymer precursor molecules according to (ii) in the presence of at least magnetic cores (M) thereby forming particles comprising at least magnetic cores (M), preferably at least two magnetic cores (M), which are embedded in a polymer matrix (P1), wherein the polymer matrix (P1) preferably comprises, more preferably consists of, a cross-linked polymer, and
(iv) (iv) preferably, hypercrosslinking the polymer matrix (P1) of the polymer particles obtained in (iii),
to obtain magnetic particles.
32. The method of embodiment 31, wherein in (iii), at least magnetic cores (M) are embedded in the matrix.
33. The method of any of embodiments 31 or 32, wherein the hypercrosslinking in (iv) is performed by a friedel-crafts reaction.
34. The method of embodiment 33, wherein the hypercrosslinking in (iv) is carried out at a temperature in the range of-30 to 120 ℃.
35. The process of any of embodiments 31 through 34, wherein the hypercrosslinking in (iv) is carried out in the presence of a catalyst comprising a Lewis acid selected from the group consisting of FeCl3、ZnCl2、AlCl3、BF3、SbCl5、SnCl4、TiCl4、SiCl4And mixtures of two or more thereof, more preferably consisting thereof; more preferably, the hypercrosslinking in (iv) is carried out in the presence of a catalyst comprising FeCl3Or ZnCl2Or a mixture thereof, more preferably consisting thereof.
36. The method of any of embodiments 31 through 35, wherein the hypercrosslinking in (iv) is performed in a solvent selected from dichloroethane, toluene, acetonitrile, dioxane, DMF, diethyl ether, THF, benzene, xylene, alkane, dichloromethane, chloroform, chlorobenzene, carbon tetrachloride, NMP, dichlorobenzene, trichlorobenzene, ether, cycloalkane, organic halide, or a mixture of two or more thereof, more preferably in a solvent selected from dichloroethane, toluene, acetonitrile, DMF, diethyl ether, THF, benzene, xylene, alkane, dichloromethane, chloroform, chlorobenzene, carbon tetrachloride, dioxane, more preferably in a solvent selected from THF, acetonitrile, DMF, dioxane, and toluene.
37. The process of any of embodiments 31 through 36 wherein the hypercrosslinking in (iv) is carried out with a crosslinking agent selected from the group consisting of dichloroethane, dichloromethane, chloroform, carbon tetrachloride, dichlorobenzene, trichlorobenzene, dichloroalkane, dibromoalkane, diiodoalkane, trichloroalkane, tribromoalkane, triiodoalkane, dimethoxymethane, dimethoxyethane, and mixtures of two or more thereof.
38. The method of any of embodiments 31 to 37, wherein said method further comprises the step of :
(v) (iv) functionalizing the surface of the polymer particles according to (iii) or (iv).
39. The method of embodiment 38, wherein in step (v), the polymer particles are functionalized with a group selected from the group consisting of: -OH, -COOH, diethylaminoEthanol, R-SO2-OH、-NH2、R-SO2-OH、-RNH、-R2N、-R3N+-CH3、-C2H5、-C4H9、-C8H17、-C18H37、-C6H5、-C6H9NO6Phenyl-hexyl, biphenyl, hydroxyapatite, boronic acid, biotin, azide, epoxide, alkyl, aryl, cycloalkyl, heteroaryl, heterocycloalkyl, amino acid, -COOR, -COR, -OR, antibodies and fragments thereof, aptamers, nucleic acids, and receptor proteins OR binding domains thereof.
40. The process of any of embodiments 31 to 39 wherein the polymerization in (iii) is suspension polymerization.
41. The process of any of embodiments 31 through 40, wherein the polymerizing in (iii) is carried out in the presence of an initiator selected from the group consisting of azobis (isobutyronitrile) (AIBN), 2 '-azobis (2-methylbutyronitrile) (VAZO 67), 1' -azobis (cyanocyclohexane) (VAZO 88), Benzoyl Peroxide (BPO), 2 '-azobis (2-amidinopropane) dihydrochloride (AAPH), and 4, 4' -azobis (4-cyanopentanoic acid) (ACVA).
42. The method of any of embodiments 31 to 41, wherein step (iii) comprises:
(iii.1) providing a composition (A) comprising a polymer precursor molecule according to (ii), at least magnetic cores (M) according to (i), at least organic solvents, at least initiators and an aqueous phase, wherein the organic solvents are immiscible with water and wherein, optionally, the water comprises at least additives and
(iii.2) stirring composition (A) to obtain emulsion (B),
wherein the emulsion is preferably an organic solvent-in-water emulsion.
43. The method of embodiment 42, wherein said at least additives are selected from the group consisting of insoluble carbonates, silicates, talc, gelatin, pectin, starch, cellulose derivatives, insoluble phosphates, PVA, salts, NaCl, KCl, and PVP.
44. The process of embodiment 42 or 43, wherein the stirring is carried out with an overhead stirrer.
45. The method of any of embodiments 29 to 43, wherein the polymerizing in (iii) is conducted in the absence of any surfactant.
46. The method of any of embodiments 31 to 45, wherein step (iii) is performed for a time in the range of 1 hour to 30 hours, preferably 1 hour to 8 hours.
47. The process of any of embodiments 31 through 46, wherein step (iii) is carried out at a temperature in the range of 0 ℃ to 100 ℃, preferably 40 ℃ to 90 ℃.
48. The method of any of embodiments 31-47, wherein step (i) comprises:
(i.1) providing at least magnetic nanoparticles, and
(i.2) coating the at least nanoparticles with a coating C1,
the coating C1 is preferably selected from the group consisting of surfactants, silicas, silicates, silanes, phosphates, phosphonates, phosphonic acids, and mixtures of two or more thereof
To obtain at least magnetic cores (M).
49. The method of embodiment 48, wherein (i) comprises:
(i.1.1) providing more than 20 magnetic nanoparticles, and preferably at least 21 nanoparticles, with at least coating C2,
(i.1.2) aggregating the more than 20 nanoparticles, thereby forming a nanoparticle, and
(i.1.3) optionally coating the superparticle according to (i.2) with at least coating layers C1,
to obtain at least magnetic cores (M).
50. The method of embodiment 49, wherein the at least coating layers C2 are selected from the group consisting of dicarboxylic acids, dicarboxylic acid salts, dicarboxylic acid derivatives, polyacrylic acids, polyacrylic acid salts, polyacrylic acid derivatives, tricarboxylic acids, tricarboxylic acid salts, tricarboxylic acid derivatives, amino acids, amino acid salts, amino acid derivatives, surfactants, salts of surfactants, fatty acids, fatty acid salts, and fatty acid derivatives.
51. A magnetic particle obtained or obtainable by the method of any of embodiments 31-50, wherein the particle is preferably a substantially spherical particle.
52. Use of the magnetic particles of any of embodiments 1-30 or the magnetic particles according to embodiment 51 for qualitative and/or quantitative determination of at least analytes in a fluid.
53. The use of embodiment 52, wherein the analyte is selected from the group consisting of a steroid, a sugar, a vitamin, a drug, an organic compound, a protein, a nucleic acid, a sugar, and a mixture of two or more thereof.
54. The use of embodiment 52 or 53, wherein the analyte is enriched by the magnetic particles of embodiments 1-30 or the magnetic particles according to embodiment 51.
55. The use of any of embodiments , wherein said analyte is determined by mass spectrometry, UV-vis, NMR, IR.
56. The use of any of embodiments , wherein the fluid is selected from the group of fluids consisting of a bodily fluid, a liquid or dissolved environmental sample, and a solution of a mixture of chemical compounds.
57. Use of the previous embodiment, wherein the magnetic particles of any of in embodiments 1-30 or the magnetic particles according to embodiment 51 reduce matrix carryover of the fluid.
58. The use of embodiment 57, wherein the qualitative and/or quantitative determination of at least analytes in said fluid relates to diagnostic purposes, substance abuse testing, environmental control, food safety, quality control, purification, or manufacturing processes.
59. A method for determining at least analytes in a fluid sample, comprising the steps of:
a) contacting the magnetic particles of the invention or the magnetic particles obtained by the method of the invention with a fluid sample comprising or suspected to comprise at least analytes, and
b) determining at least analytes bound to the magnetic particles.
60. The method of embodiment 59, wherein step a) of said method is performed for a time and under conditions sufficient to allow said at least analytes to bind to said magnetic particles.
61. The method of
a1) preferably without eluting the at least analytes, to which at least moieties of at least analytes are bound, and/or
a2) Eluting the at least analytes from the magnetic particles under conditions suitable to allow elution of the at least analytes.
62. The method of any of embodiments 59 through 61, wherein the determining comprises determining the presence or absence of bound analyte on the magnetic particles or determining the amount of analyte bound to the magnetic particles.
Further optional features and embodiments of the invention will be disclosed in more detail in the subsequent description of preferred embodiments, preferably in combination with the dependent claims. Wherein the respective optional features may be realized in an independent manner as well as in any feasible combination, as will be appreciated by the person skilled in the art. The scope of the invention is not limited by the preferred embodiments. Embodiments are schematically depicted in the drawings. In the drawings, like reference numbers indicate identical or functionally equivalent elements.
All references cited in this specification are incorporated herein by reference to the specifically mentioned disclosure as well as in their entirety.
Drawings
FIG. 1 is a schematic representation of a magnetic particle comprising magnetic cores (M) (10a + 10b) having 1-20 nanoparticles (10a) and a coating C1(10b) and a polymer matrix (P) (11)
FIG. 2 is a schematic representation of a more preferred magnetic particle comprising more than , i.e. three magnetic cores (M) (10a + 10b) each having 1-20 nanoparticles (10a) and a coating C1(10b), and a polymer matrix (P) (11)
FIG. 3 is a schematic view of a magnetic particle having a polymer matrix (P) (11) and magnetic cores (M) consisting of a super-particle formed by aggregating a plurality of nanoparticles (10a) coated with a coating C2(10C), the magnetic cores (M) further having a coating C1(10b)
FIG. 4 is a schematic illustration of a more preferred magnetic particle having a polymer matrix (P) (11) and more than magnetic cores (M) consisting of a super-particle formed by aggregating a plurality of nanoparticles (10a) coated with a coating C2(10C), said magnetic cores (M) further having a coating C1(10b)
Fig. 5 shows the calculated saturation magnetization of particles of different sizes synthesized by Xu et al.
Fig. 6 shows an enrichment workflow for sample analysis using beads as described herein.
Examples
The following examples are intended only to illustrate the invention. They should not be construed as limiting the scope of the invention in any way.
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