Anti-mullerian hormone antibodies, reagents and kits for detecting mullerian hormone

Abstract

The application discloses an antibody against Mullerian hormone or a reagent and kit for detecting Mullerian hormone, and relates to the field of antibodies. The anti-Mullerian hormone antibody disclosed by the application comprises a heavy chain complementarity determining region and a light chain complementarity determining region, the antibody provides an important raw material source for detection of Mullerian hormone, and has good affinity and activity.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of antibody technology, in particular to an anti-Mullerian hormone antibody, a reagent for detecting Mullerian hormone and a kit. BACKGROUND

[0002] Anti-Mullerian hormone (AMH) is a member of the transforming growth factor beta superfamily, which was first discovered by Professor Alfred Jost in 1974. It is a dimeric glycoprotein composed of two 70kD protein subunits through disulfide bonds, with a relative molecular mass of 140kD, and its main function is to affect the growth and differentiation of follicles. The gene encoding AMH in humans is located on chromosome 19p13.3, with a size of 2.4-2.8kb and containing 5 exons. Studies have shown that the N-terminal region of AMH plays a key role in maintaining the integrity of the protein and its activity. During metabolic transport, a specific site (amino acid 451) of AMH can be cut to form a pro-fragment (pro-AMH, 26-451aar) and a mature fragment (Mature AMH, 452-560aar), and the two fragments can be connected together through non-covalent bonds.

[0003] AMH plays an important role in the development of gonadal organs, and is one of the important markers of male and female gonadal function. In the sertoli cells of male fetuses, transcription factor SOX-9 (SOX9) can activate AMH to combine with anti-Mullerian hormone receptor type II (AMHR2) and cause the Mullerian duct in the male embryo to degenerate, thereby preventing the formation of the fallopian tube, uterus and upper part of the vagina, and allowing the male reproductive tract to develop normally. In female embryos, the Mullerian duct will differentiate into uterine tubes and fallopian tubes. AMH is also a product of the preantral follicle and small antral follicle granulosa cells in women. Since puberty, the level of AMH in the serum slowly decreases over time, and decreases to an undetectable level by ELISA during menopause. The normal value of AMH is between 2-6.8 ng / ml, and the higher the value, the more abundant the egg stock, the longer the golden period suitable for pregnancy, and the lower the value, the poorer the ovarian function. After the age of 35, the AMH value will begin to decrease rapidly, and when it is lower than 0.7 ng / ml, it indicates that the egg stock is severely insufficient and almost impossible to get pregnant. If the value is greater than 6.8, it can be considered that the person has a polycystic ovary syndrome. When ovulation needles and drugs are used, the ovary is also prone to overreaction and the discharge of too many eggs, causing ovarian hyperstimulation syndrome. Therefore, the level of AMH can be used to judge the egg stock and ovarian function, and can be used as a diagnostic basis for ovarian response, polycystic ovary syndrome (PCOS), and ovarian hyperstimulation syndrome (OHSS) in in vitro fertilization combined with embryo transfer (IVF).

[0004] Currently, the commonly used methods for detecting AMH in clinical practice at home and abroad include enzyme-linked immunosorbent assay (ELISA), electrochemiluminescence method, and chemiluminescence method. Among them, the ELISA method is time-consuming, low in sensitivity, high in background value, prone to false positive and false negative results, and requires professional operation. The electrochemiluminescence method and the chemiluminescence method, such as the ELECSYS AMH detector of Roche, are gradually replacing the ELISA method, but are expensive and not suitable for single-person and small-batch detection. In addition, special instrument operators are required, and the maintenance and detection costs are high, which is not suitable for wide use.

[0005] The above methods all need to use anti-AMH monoclonal antibodies, and therefore, there is a strong demand in the art for anti-Mullerian hormone antibodies with good activity and affinity. SUMMARY

[0006] The present application provides an antibody or a functional fragment thereof, which provides an important raw material source for the detection of Mullerian hormone and has good activity and affinity.

[0007] In order to achieve the above object, according to an aspect of the present application, there is provided an antibody or a functional fragment thereof, comprising HCDR1, HCDR2, HCDR3 and LCDR1, LCDR2, LCDR3, the HCDR1, HCDR2, HCDR3 being an amino acid sequence identical to HCDR1, HCDR2, HCDR3 of a heavy chain variable region shown in any one of SEQ ID NO: 17, 18; the LCDR1, LCDR2, LCDR3 being an amino acid sequence identical to LCDR1, LCDR2, LCDR3 of a light chain variable region shown in any one of SEQ ID NO: 19, 20, 21, 22.

[0008] In order to achieve the above object, according to a second aspect of the present application, there is provided an antibody or a functional fragment thereof, comprising a heavy chain variable region and / or a light chain variable region, the amino acid sequence of the heavy chain variable region being shown in any one of SEQ ID NO: 17, 18; the amino acid sequence of the light chain variable region being shown in any one of SEQ ID NO: 19, 20, 21, 22.

[0009] In order to achieve the above object, according to a third aspect of the present application, there is provided an antibody or a functional fragment thereof, comprising a heavy chain and / or a light chain, the amino acid sequence of the heavy chain being shown in any one of SEQ ID NO: 23, 24; the amino acid sequence of the light chain being shown in any one of SEQ ID NO: 25, 26, 27, 28.

[0010] In order to achieve the above object, according to a fourth aspect of the present application, there is provided an antibody conjugate, comprising the above-mentioned antibody or functional fragment thereof.

[0011] In order to achieve the above object, according to a fifth aspect of the present application, there is provided a reagent or kit, comprising the above-mentioned antibody or functional fragment thereof or the above-mentioned antibody conjugate.

[0012] In order to achieve the above object, according to a sixth aspect of the present application, there is provided a method for detecting Mullerian duct hormone, comprising: a) contacting the above-mentioned antibody or functional fragment thereof, antibody conjugate, or reagent or kit with Mullerian duct hormone in a sample to be detected under conditions sufficient for an antibody / antigen binding reaction to occur, to form an immunocomplex; and b) detecting the presence of the immunocomplex, the presence of the complex indicating the presence of the antigen in the test sample.

[0013] In order to achieve the above object, according to a seventh aspect of the present application, there is provided a nucleic acid, encoding the above-mentioned antibody or functional fragment thereof.

[0014] To achieve the above object, according to an eighth aspect of the present application, there is provided a vector comprising the above-mentioned nucleic acid.

[0015] To achieve the above object, according to a ninth aspect of the present application, there is provided a cell comprising the above-mentioned nucleic acid, vector or expressing the above-mentioned antibody or functional fragment thereof.

[0016] To achieve the above object, according to a tenth aspect of the present application, there is provided a method for preparing the above-mentioned antibody or functional fragment thereof, comprising culturing the above-mentioned cell.

[0017] To achieve the above object, according to an eleventh aspect of the present application, there is provided a use of the above-mentioned antibody or functional fragment thereof, antibody conjugate, reagent or kit in detecting or preparing a product for detecting Mullerian Inhibiting Substance. BRIEF DESCRIPTION OF DRAWINGS

[0018] To more clearly illustrate the technical solutions of the embodiments of the present application, the following will briefly introduce the drawings needed to be used in the embodiments. It should be understood that the following drawings only show some of the embodiments of the present application, and therefore should not be considered as a limitation to the scope. For those skilled in the art, other related drawings can also be obtained without creative labor on the basis of these drawings.

[0019] Figure 1 Results of reducing SDS-PAGE for Anti-AMH 15G10Rmb1. DETAILED DESCRIPTION

[0020] In a first aspect, the embodiments of the present application provide an antibody or functional fragment thereof, comprising HCDR1, HCDR2, HCDR3 and LCDR1, LCDR2, LCDR3, the HCDR1, HCDR2, HCDR3 are identical to the HCDR1, HCDR2, HCDR3 of the heavy chain variable region shown in any one of SEQ ID NO: 17, 18; the LCDR1, LCDR2, LCDR3 are identical to the LCDR1, LCDR2, LCDR3 of the light chain variable region shown in any one of SEQ ID NO: 19, 20, 21, 22.

[0021] It is to be noted that the HCDR1, HCDR2 and HCDR3 are the amino acid sequences identical to the HCDR1, HCDR2, HCDR3 of the same heavy chain variable region defined in the antibody or functional fragment thereof of the first aspect, and the LCDR1, LCDR2 and LCDR3 are the amino acid sequences identical to the LCDR1, LCDR2, LCDR3 of the same light chain variable region defined in the antibody or functional fragment thereof of the first aspect.

[0022] For example, the HCDR1, HCDR2, HCDR3 are the amino acid sequences identical to the HCDR1, HCDR2, HCDR3 of the heavy chain variable region shown in SEQ ID NO: 17, and the LCDR1, LCDR2, LCDR3 are the amino acid sequences identical to the LCDR1, LCDR2, LCDR3 of the light chain variable region shown in SEQ ID NO: 19.

[0023] Under the Kabat definition, the amino acid sequences of the HCDR1, HCDR2, HCDR3 of the heavy chain variable region SEQ ID NO: 17 are shown in SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, respectively, and the amino acid sequences of the HCDR1, HCDR2, HCDR3 of the antibody or functional fragment thereof are also shown in SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, respectively.

[0024] Under the Kabat definition, the amino acid sequences of the LCDR1, LCDR2, LCDR3 of the light chain variable region SEQ ID NO: 19 are shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, respectively, and the amino acid sequences of the LCDR1, LCDR2, LCDR3 of the antibody or functional fragment thereof are also shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, respectively.

[0025] In the present application, the term "antibody" is used in the broadest sense, and can include full-length monoclonal antibodies, bispecific or multispecific antibodies, and chimeric antibodies, as long as they exhibit the desired biological activity.

[0026] In the present application, the term "complementarity determining region", "CDR" or "CDRs" refers to the highly variable regions of the heavy and light chains of immunoglobulins, and refers to the regions that contain one or more, or even all, of the amino acid residues that play a major role in binding of the antibody or antigen-binding fragment to its recognized antigen or epitope. In the detailed description of the present application, the CDRs refer to the highly variable regions of the heavy and light chains of the antibody.

[0027] In the present disclosure, heavy chain complementarity determining regions are denoted as HCDR, which includes HCDR1, HCDR2 and HCDR3; light chain complementarity determining regions are denoted as LCDR, which includes LCDR1, LCDR2 and LCDR3.

[0028] Methods for defining CDRs are well known in the art and include the Kabat definition, the Chothia definition, the IMGT definition, the Contact definition and the AbM definition. As used herein, the "Kabat definition" refers to the definition system described in Kabat et al., U.S. Dept. of Health and Human Services, "Sequence of Proteins of Immunological Interest" (1983). The "Chothia definition" is described in Chothia et al., J Mol Biol 196:901-917 (1987). Other methods of defining CDRs can not strictly follow one of the above schemes but will still overlap at least a portion of the CDR regions defined by Kabat, although they can be shortened or lengthened according to prediction or experimental results for particular residues or groups of residues. Exemplary defined CDRs are listed in Table 1 below, with slight variations in nomenclature in different references. Given the variable region amino acid sequence of a given antibody, one of ordinary skill in the art can routinely determine which residues comprise particular CDRs. It is noted that CDRs defined by other methods than those in Table 1 are also within the scope of the present disclosure.

[0029] Table 1: CDR definitions 1

[0030] CDR Kabat AbM 2 ]] AbM IMGT Chothia [H31-H35 3 ]]> H26-H35 3 ]] [H26~H33..5 5 ]]> H26-H32..34 4 ]]> HCDR1 H31-H35 H26-H35 H26-H33 H26-H32 HCDR2 H50-H65 H50-H58 H51-H57 H52-H56 HCDR3 H95-H102 H95-H102 H93-H102 H95-H102 LCDR1 L24-L34 L24-L34 L27-L32 L24-L34 LCDR2 L50-L56 L50-L56 L50-L51 L50-L56

[0031] 1 The numbering of all CDR definitions in Table 1 is according to the Kabat numbering system (see below), with "H+ numbers" for amino acid positions on the heavy chain and "L+ numbers" for amino acid positions on the light chain. One of ordinary skill in the art can unambiguously correlate this Kabat numbering system to any variable region sequence without reliance on any experimental data other than the sequence itself. As used herein, the "Kabat numbering" refers to the numbering system described in Kabat et al., U.S. Dept. of Health and Human Services, "Sequence of Proteins of Immunological Interest" (1983).

[0032] 2"AbM" as used in Table 1, with a lower case "b", refers to CDRs defined by the "AbM" antibody modeling software of Oxford Molecular.

[0033] 3 If neither H35A nor H35B is present, then CDR-H1 ends at position 35; if only H35A is present, then CDR-H1 ends at position 35A; if both H35A and H35B are present, then CDR-H1 ends at position 35B.

[0034] 4 If neither H35A nor H35B is present, then CDR-H1 ends at position 32; if only H35A is present, then CDR-H1 ends at position 33; if both H35A and H35B are present, then CDR-H1 ends at position 34.

[0035] 5 If neither H35A nor H35B is present, then CDR-H1 ends at position 33; if only H35A is present, then CDR-H1 ends at position 34; if both H35A and H35B are present, then CDR-H1 ends at position 35.

[0036] According to embodiments of the application, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, or LCDR3 is defined by any one of the Kabat, Chothia, IMGT, AbM, or Contact system, or a combination of more than one of the systems.

[0037] In some optional embodiments of the application, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are defined by the Kabat system.

[0038] In some optional embodiments of the application, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are defined by the Chothia system.

[0039] In some optional embodiments of the application, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are defined by the IMGT system.

[0040] In some optional embodiments of the application, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 are defined by the AbM system.

[0041] In some embodiments of the application, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the Contact system.

[0042] In some embodiments of the application, the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the Kabat, Chothia, IMGT, AbM or Contact system.

[0043] According to embodiments of the application, the Kabat, Chothia, AbM or IMGT system defined HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 or LCDR3 amino acid sequence corresponding Kabat numbered positions are as follows:

[0044] LCDR3 L89-L97 L89-L97 L89-L97 L89-L97 Combination Heavy chain Light chain SEQ ID NO: 23 SEQ ID NO: 25 SEQ ID NO: 24 SEQ ID NO: 26 SEQ ID NO: 23 SEQ ID NO: 26 SEQ ID NO: 23 SEQ ID NO: 27 SEQ ID NO: 23 SEQ ID NO: 28 Antibody name Heavy chain Light chain Anti-AMH 15G10Rmb1 SEQ ID NO: 23 SEQ ID NO: 25 Anti-AMH 15G10Rmb2 SEQ ID NO: 24 SEQ ID NO: 26 Anti-AMH 15G10Rmb3 SEQ ID NO: 23 SEQ ID NO: 26 Anti-AMH 15G10Rmb4 SEQ ID NO: 23 SEQ ID NO: 27 Anti-AMH 15G10Rmb5 SEQ ID NO: 23

[0045] According to embodiments of the application, the HCDRs and LCDRs are defined by the Kabat system.

[0046] In alternative embodiments, the embodiments of the application provide an antibody or functional fragment thereof, the antibody or functional fragment thereof comprising the following complementarity determining regions:

[0047] HCDR1 comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.

[0048] HCDR2 comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 2.

[0049] HCDR3 comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 3.

[0050] LCDR1 comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 4 or 30.

[0051] LCDR2 comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 5.

[0052] LCDR3 comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 6 or 31.

[0053] In the present application, the "framework region" or "FR" region, which includes the heavy chain framework region and the light chain framework region, refers to the region of the antibody heavy chain variable region and the light chain variable region other than the CDR; wherein the heavy chain framework region can be further subdivided into adjacent regions separated by CDRs, comprising HFR1, HFR2, HFR3 and HFR4 framework regions; the light chain framework region can be further subdivided into adjacent regions separated by CDRs, comprising LFR1, LFR2, LFR3 and LFR4 framework regions.

[0054] In the present application, the heavy chain variable region is obtained by connecting the following numbered CDRs and FRs in the following combination: HFR1-HCDR1-HFR2-HCDR2-HFR3-HCDR3-HFR4; the light chain variable region is obtained by connecting the following numbered CDRs and FRs in the following combination: LFR1-LCDR1-LFR2-LCDR2-LFR3-LCDR3-LFR4.

[0055] In an alternative embodiment, the antibody or functional fragment thereof further has at least one of HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3 and LFR4;

[0056] The HFR1 comprises / like SEQ ID NO: 7 or an amino acid sequence having at least 80% identity thereto;

[0057] The HFR2 comprises / like SEQ ID NO: 8 or an amino acid sequence having at least 80% identity thereto;

[0058] The HFR3 comprises / like SEQ ID NO: 9 or an amino acid sequence having at least 80% identity thereto;

[0059] The HFR4 comprises / like SEQ ID NO: 10 or an amino acid sequence having at least 80% identity thereto;

[0060] The LFR1 comprises / like SEQ ID NO: 11 or an amino acid sequence having at least 80% identity thereto;

[0061] The LFR2 comprises / like SEQ ID NO: 12 or an amino acid sequence having at least 80% identity thereto;

[0062] The LFR3 comprises / like SEQ ID NO: 13 or an amino acid sequence having at least 80% identity thereto;

[0063] The LFR4 comprises / like SEQ ID NO: 14 or an amino acid sequence having at least 80% identity thereto;

[0064] It should be noted that in other embodiments, each of the framework regions of the antibody or functional fragment thereof provided by the present application can have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the corresponding framework region (SEQ ID NO: 7, 8, 9, 10, 11, 12, 13, or 14) described above.

[0065] In an alternative embodiment, the HFR3 comprises / has the amino acid sequence set forth in SEQ ID NO: 9 or 29.

[0066] In an alternative embodiment, the antibody or functional fragment thereof binds Mullerian Inhibiting Hormone with an affinity of KD< 7.21 x 10 -7 M.

[0067] In an alternative embodiment, the antibody or functional fragment thereof binds Mullerian Inhibiting Hormone with an affinity of KD≤ 10 -7 M. -8 M. - 9 M. -10 M. -11 M. -12 M.

[0068] In an alternative embodiment, the antibody or functional fragment thereof binds Mullerian Inhibiting Hormone with an affinity of KD≤ 3.52 x 10 -8 M.

[0069] There are many methods for determining the affinity (KD) of an antibody, which can be classified into thermodynamic detection methods, kinetic detection methods, and dynamic equilibrium detection methods according to the detection principle. Among them, the thermodynamic detection method is commonly known as isothermal titration calorimetry (ITC); the kinetic detection method is commonly known as surface plasmon resonance (SPR) and bio-layer interferometry (BLI); and the dynamic equilibrium detection method is commonly known as enzyme-linked immunosorbent assay (ELISA) and the like.

[0070] In an alternative embodiment, the determination of KD is performed using a kinetic detection method; alternatively, surface plasmon resonance, for example, by using a biosensor system such as a Biacore® system. In an alternative embodiment, the determination of KD is performed using a kinetic detection method; alternatively, surface plasmon resonance, for example, by using a biosensor system such as a Biacore® system.

[0071] In a second aspect, the embodiments of the present application provide an antibody or a functional fragment thereof, comprising a heavy chain variable region and / or a light chain variable region, wherein the amino acid sequence of the heavy chain variable region is shown in any one of SEQ ID NO: 17 or 18, and the amino acid sequence of the light chain variable region is shown in any one of SEQ ID NO: 19, 20, 21 or 22.

[0072] In an alternative embodiment, the antibody or the functional fragment thereof of the first aspect and the second aspect comprises any one of the following combinations of the heavy chain variable region and the light chain variable region:

[0073]

[0074]

[0075] In an alternative embodiment, the antibody or the functional fragment thereof further comprises a constant region.

[0076] In an alternative embodiment, the constant region comprises a heavy chain constant region and / or a light chain constant region.

[0077] In an alternative embodiment, the heavy chain constant region is selected from the heavy chain constant region of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE or IgD, and the light chain constant region is selected from the light chain constant region of kappa type or lambda type.

[0078] In an alternative embodiment, the species origin of the constant region is bovine, equine, bovine, porcine, ovine, rat, mouse, dog, cat, rabbit, donkey, deer, mink, chicken, duck, goose, turkey, cock or human.

[0079] In an alternative embodiment, the species origin of the constant region is mouse.

[0080] In an alternative embodiment, the sequence of the heavy chain constant region (CH) is shown in SEQ ID NO: 15, and the sequence of the light chain constant region (CL) is shown in SEQ ID NO: 16.

[0081] It should be noted that in other embodiments, the sequence of the constant region can have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the above constant region (SEQ ID NO: 15 or 16).

[0082] In an alternative embodiment, the functional fragment is selected from any one of F(ab’)2, Fab’, Fab, Fv and scFv of the antibody.

[0083] The functional fragment of the antibody described above usually has the same binding specificity as the antibody from which it is derived. It is easy for those skilled in the art to understand that the functional fragment of the antibody described above can be obtained by methods such as enzymatic digestion (including pepsin or papain) and / or by methods of splitting disulfide bonds by chemical reduction, based on the structural basis of the complete antibody disclosed in the present application. Based on the structural basis of the complete antibody disclosed in the present application, those skilled in the art can easily obtain the functional fragment described above.

[0084] The functional fragment of the antibody described above can also be obtained by recombinant genetic engineering techniques also known to those skilled in the art or by, for example, an automatic peptide synthesizer, such as an automatic peptide synthesizer sold by Applied BioSystems and the like.

[0085] In a third aspect, the present application provides an antibody or a functional fragment thereof, comprising a heavy chain and / or a light chain, wherein the amino acid sequence of the heavy chain is shown in any one of SEQ ID NO: 23, 24, and the amino acid sequence of the light chain is shown in any one of SEQ ID NO: 25, 26, 27, 28.

[0086] In an alternative embodiment, the antibody described in the first aspect, the second aspect, and the third aspect of the present application comprises a heavy chain and a light chain in any one of the following combinations:

[0087] SEQ ID NO: 28 Concentration (ng / ml) Control 1 Anti-AMH 15G10Rmb1 Anti-AMH 15G10Rmb2 2 Anti-AMH 15G10Rmb3 Anti-AMH 15G10Rmb4 3 Anti-AMH 15G10Rmb5 Sample concentration (ng / ml) 4 4°C, 21-day sample -80°C, 21-day sample 5 37°C, 21-day sample 4°C, 21-day sample

[0088] In a fourth aspect, the present application provides an antibody conjugate, comprising the antibody or the functional fragment thereof described above.

[0089] In an alternative embodiment, the antibody conjugate described above further comprises biotin or a biotin derivative conjugated to the antibody or the functional fragment thereof.

[0090] In an alternative embodiment, the antibody conjugate described above further comprises a label conjugated to the antibody or the functional fragment thereof.

[0091] In an alternative embodiment, the label described above refers to a kind of substance having a property that can be directly observed by the naked eye or detected or probed by an instrument, such as luminescence, color development, radioactivity, and the like, through which qualitative or quantitative detection of the corresponding target can be achieved.

[0092] In an alternative embodiment, the label described above includes but is not limited to fluorescent dyes, enzymes, radioisotopes, chemiluminescent reagents, and nanoparticle-based labels.

[0093] In actual use, those skilled in the art can select appropriate labels according to the detection conditions or actual needs, and regardless of the label used, it falls within the scope of the present application.

[0094] In optional embodiments, the fluorescent dye includes, but is not limited to, fluorescein-based dyes and their derivatives (such as, but not limited to, fluorescein isothiocyanate (FITC), hydroxycoumarin (FAM), tetra-chloro-fluorescein (TET), and the like or analogs thereof), rhodamine-based dyes and their derivatives (such as, but not limited to, red rhodamine (RBITC), tetramethyl rhodamine (TAMRA), rhodamine B (TRITC), and the like or analogs thereof), Cy series dyes and their derivatives (such as, but not limited to, Cy2, Cy3, Cy3B, Cy3.5, Cy5, Cy5.5, Cy3, and the like or analogs thereof), Alexa series dyes and their derivatives (such as, but not limited to, Alexa Fluor 350, 405, 430, 488, 532, 546, 555, 568, 594, 610, 33, 647, 680, 700, 750, and the like or analogs thereof), and protein-based dyes and their derivatives (such as, but not limited to, phycoerythrin (PE), phycocyanin (PC), allophycocyanin (APC), peridinin-chlorophyll protein (preCP), and the like).

[0095] In optional embodiments, the enzyme includes, but is not limited to, horseradish peroxidase, alkaline phosphatase, beta-galactosidase, glucose oxidase, carbonic anhydrase, acetylcholinesterase, and 6-phosphogluconate dehydrogenase.

[0096] In optional embodiments, the radioisotope includes, but is not limited to, 212Bi, 131I, 111In, 90Y, 186Re, 211At, 125I, 188Re, 153Sm, 213Bi, 32P, 94mTc, 99mTc, 203Pb, 67Ga, 68Ga, 43Sc, 47Sc, 110mIn, 97Ru, 62Cu, 64Cu, 67Cu, 68Cu, 86Y, 88Y, 121Sn, 161Tb, 166Ho, 105Rh, 177Lu, 172Lu, and 18F.

[0097] In optional embodiments, the chemiluminescent reagent includes, but is not limited to, luminol and its derivatives, lucigenin, crustacyanin, ruthenium bis-2,2'-bipyridine and its derivatives, acridinium ester and its derivatives, dioxetane and its derivatives, luminol and its derivatives, and peroxyoxalate and its derivatives.

[0098] In optional embodiments, the nanoparticle-based label includes, but is not limited to, nanoparticles, colloids, organic nanoparticles, magnetic nanoparticles, quantum dot nanoparticles, and rare earth complex nanoparticles.

[0099] In optional embodiments, the colloid includes, but is not limited to, colloidal metals, dispersed dyes, dye-labeled microspheres, and latex.

[0100] In optional embodiments, the colloidal metal includes, but is not limited to, colloidal gold, colloidal silver and colloidal selenium.

[0101] In optional embodiments, the colloidal metal is colloidal gold.

[0102] In optional embodiments, the antibody conjugate further comprises a solid support coupled to the antibody or functional fragment thereof.

[0103] In optional embodiments, the solid support is selected from the group consisting of microspheres, plates and membranes.

[0104] In optional embodiments, the solid support includes, but is not limited to, magnetic microspheres, plastic microspheres, plastic microparticles, microwell plates, glass, capillary tubes, nylon and nitrocellulose membranes.

[0105] In a fifth aspect, the present application provides a reagent or kit comprising the antibody or functional fragment thereof or the antibody conjugate as described above.

[0106] As described above, the antibody or functional fragment thereof in some embodiments or examples of the present application can effectively bind to the Mullerian duct hormone, and thus the reagent or kit comprising the Mullerian duct hormone antibody or functional fragment thereof can effectively detect the Mullerian duct hormone qualitatively or quantitatively. The reagent or kit provided by the present application can be used in, for example, immunoblotting, immunoprecipitation and other tests involving the specific binding of the Mullerian duct hormone and its antibody. As described above, the antibody or functional fragment thereof in some embodiments or examples of the present application has higher binding activity or affinity to the Mullerian duct hormone, and thus the reagent or kit comprising the antibody or functional fragment thereof has higher detection sensitivity or specificity.

[0107] In a sixth aspect, the present application provides a method for detecting the Mullerian duct hormone, comprising: a) contacting the antibody or functional fragment thereof, the antibody conjugate, the reagent or kit as described above with the Mullerian duct hormone in a sample to be tested under conditions sufficient to allow the antibody / antigen binding reaction to occur to form an immunocomplex; and b) detecting the presence of the immunocomplex, wherein the presence of the complex indicates the presence of the antigen in the test sample.

[0108] In optional embodiments, the immunocomplex further comprises a second antibody that binds to the antibody or functional fragment thereof.

[0109] In optional embodiments, the immunocomplex further comprises a second antibody that binds to the Mullerian duct hormone.

[0110] In a seventh aspect, the present application provides a nucleic acid molecule encoding the antibody or functional fragment thereof as described above.

[0111] In an eighth aspect, the present application provides a vector comprising the nucleic acid molecule described above.

[0112] In a ninth aspect, the present application provides a cell comprising the vector described above.

[0113] In a tenth aspect, the present application provides a method for preparing an antibody or a functional fragment thereof, comprising culturing the cell described above.

[0114] In an eleventh aspect, the present application provides use of the antibody or a functional fragment thereof, an antibody conjugate, or the reagent or kit described above in detecting Mullerian duct hormone or in preparing a product for detecting Mullerian duct hormone.

[0115] On the basis of the amino acid sequence of the antibody or a functional fragment thereof disclosed in the present application, it is easy for those skilled in the art to conceive of preparing the antibody or a functional fragment thereof by using genetic engineering technology or other technologies (chemical synthesis, recombinant expression), for example, by isolating and purifying the antibody or a functional fragment thereof from the culture product of recombinant cells capable of recombinantly expressing the antibody or a functional fragment thereof as described in any one of the above, which is easy for those skilled in the art to achieve. Therefore, no matter what technology is used to prepare the antibody or a functional fragment thereof of the present application, it all falls within the protection scope of the present application.

[0116] In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below. Unless otherwise specified, the specific conditions in the embodiments are carried out according to conventional conditions or manufacturer's recommended conditions. Unless otherwise specified, the reagents or instruments used are all conventional products that can be purchased on the market.

[0117] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the preparations or unit doses herein, some methods and materials are now described. The techniques employed or contemplated herein are standard methodologies unless otherwise indicated. Materials, methods, and examples are illustrative only and not limiting.

[0118] Unless otherwise indicated, the practice of the present application will employ, unless otherwise indicated, conventional techniques of cell biology, molecular biology (including recombinant techniques), microbiology, biochemistry and immunology, which are within the skill of the art. Such techniques are explained fully in the literature, such as, Molecular Cloning: A Laboratory Manual, Second Edition (Sambrook et al., 1989); Oligonucleotide Synthesis (M. J. Gait, ed., 1984); Animal Cell Culture (R. I. Freshney, ed., 1987); Methods in Enzymology (Academic Press, Inc.); Handbook of Experimental Immunology (D. M. Weir and C. C. Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (J. M. Miller and M. P. Calos, eds., 1987); Current Protocols in Molecular Biology (F. M. Ausubel et al., eds., 1987); PCR: The Polymerase Chain Reaction (Mullis et al., eds., 1994); and Current Protocols in Immunology (J. E. Coligan et al., eds., 1991), each of which is incorporated herein by reference in its entirety.

[0119] The features and nature of the present application will be further described in connection with the following examples.

[0120] Example 1 Preparation of Anti-AMH 15G10 Monoclonal Antibody

[0121] Restriction enzymes and Prime Star DNA polymerase used in this example were purchased from Takara. MagExtractor-RNA extraction kit was purchased from TOYOBO. BD SMART TMRACE cDNA Amplification Kit was purchased from Takara Company. pMD-18T vector was purchased from Takara Company. Plasmid extraction kit was purchased from Tiangen Company. Primer synthesis and gene sequencing were completed by Invitrogen Company. Hybridoma cell strain secreting Anti-AMH 15G10 monoclonal antibody was an existing hybridoma cell strain, which was recovered for use.

[0122] (1) Antibody gene preparation

[0123] The mRNA was extracted from the hybridoma cell strain secreting Anti-AMH 15G10 monoclonal antibody, and the DNA product was obtained by RT-PCR method. After A reaction with rTaq DNA polymerase, the product was inserted into the pMD-18T vector, transformed into DH5α competent cells, and after the colonies were grown, the Heavy Chain and Light Chain gene clones were taken respectively, and each 4 clones were sent to the gene sequencing company for sequencing.

[0124] (2) Sequence analysis of Anti-AMH 15G10 antibody variable region gene

[0125] The gene sequence obtained by the above sequencing was placed in the kabat antibody database for analysis, and the VNTI11.5 software was used for analysis to determine that the amplified genes by the heavy chain and light chain primer pairs were correct. Among them, the VL gene sequence in the Light Chain amplified gene fragment was 336 bp, and there was a 57 bp leader peptide sequence in front of it; the VH gene sequence in the Heavy Chain amplified gene fragment was 354 bp, belonging to the VH1 gene family, and there was a 57 bp leader peptide sequence in front of it.

[0126] (3) Construction of recombinant antibody expression plasmid

[0127] pcDNA TM 3.4 vector is a constructed recombinant antibody eukaryotic expression vector, which has introduced HindIII, BamHI, EcoRI and other multiple enzyme digestion sites, and is named pcDNA3.4A expression vector, hereinafter referred to as 3.4A expression vector; according to the antibody variable region gene sequencing results in the above pMD-18T, the VL and VH gene specific primers of the antibody were designed, which had HindIII, EcoRI enzyme digestion sites and protection bases at both ends, respectively. The 0.73 kb Light Chain gene fragment and the 1.44 kb Heavy Chain gene fragment were amplified by PCR amplification method.

[0128] The Heavy Chain and Light Chain gene fragments were digested with HindIII / EcoRI, and the 3.4A vector was also digested with HindIII / EcoRI. After purification and recovery of the fragments and vector, the Heavy Chain gene and Light Chain gene were ligated into the 3.4A expression vector to obtain recombinant expression plasmids of Heavy Chain and Light Chain, respectively.

[0129] 2. Recombinant antibody production

[0130] HEK293 cells were revived early and passaged to a 200ml volume to achieve a cell density of 3–5 × 10⁻⁶ cells / mL. 6 Cell density reached the required antibody concentration and cell viability >95%; cells were washed by centrifugation, reconstituted with culture medium, and the cell density was adjusted to 2.9 × 10⁻⁶ cells / ml. 6 Cells were washed at a concentration of cells / ml and reconstituted with culture medium, which served as a cell dilution buffer. Plasmid DNA and transfection reagent dilution buffers were prepared separately using culture medium. The transfection reagent dilution buffer was added to the plasmid DNA dilution buffer, mixed well, and incubated at room temperature for 15 min. This mixture was then slowly added to the cell dilution buffer over 1 min, mixed well, and samples were taken for cell counting. Cell viability after transfection was recorded and observed. The cells were then incubated at 35°C with a rotation speed of 120 rpm and a CO2 concentration of 8%. After 13 days, the samples were centrifuged and collected. The supernatant was purified using a protein A affinity chromatography column. 6 μg of the purified antibody was subjected to reducing SDS-PAGE, as shown in the figure. The reducing SDS-PAGE showed two bands: one with a Mr of 50 kDa (heavy chain) and the other with a Mr of 28 kDa (light chain).

[0131] The resulting antibody was named Anti-AMH 15G10Rmb1. Mutations were performed on Anti-AMH 15G10Rmb1 to obtain a mutant antibody. The heavy chain (H) and light chain (L) sequences of the above antibody are shown in the table below:

[0132] Table 2 Antibody Sequences

[0133] -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample

[0134] Example 2: Antibody Performance Detection

[0135] 1. Affinity Analysis

[0136] Purified antibodies were diluted in advance, and AMH antigen (purchased from Fepure Biotech) was diluted in gradient; the dissociation curve of antigen-antibody binding was tested on a Biacore 8K+ device using a CM5 chip pre-coupled with goat anti-mouse IgG, and the affinity constant, binding rate and dissociation rate were obtained by automatic fitting of the instrument.

[0137] Table 3 affinity data

[0138]

[0139]

[0140] 2. Activity identification

[0141] Coating solution (main component NaHCO3) diluted AMH antigen (purchased from Fepure Biotech) to 3ug / ml, 100uL per well, 4℃ overnight; the next day, wash solution (main components Na2HPO4+Nacl) was washed twice, and then dried; blocking solution (20% BSA+80% PBS) was added, 120uL per well, 37℃, 1h, and then dried; diluted purified antibodies and control antibodies were added, 100uL per well, 37℃, 30min; wash solution was washed 5 times, and then dried; goat anti-mouse IgG-HRP was added, 100uL per well, 37℃, 30min; wash solution was washed 5 times, and then dried; color developing solution A (50uL per well) was added, color developing solution B (50uL per well) was added, 10min; stop solution was added, 50uL per well; OD value was read on a microplate reader at 450nm (reference 630nm).

[0142] Note: A solution (main components citric acid+ sodium acetate+ acetanilide+ urea peroxide); B solution (main components citric acid+ EDTA·2Na+ TMB+ concentrated HCL); stop solution (EDTA·2Na+ concentrated H2SO4)

[0143] Table 4 activity data

[0144] -80°C, 21-day sample 93.750 46.875 23.438 11.719 5.859 0.000 37°C, 21-day sample 1.189 0.801 0.327 0.176 0.018 0.011 4°C, 21-day sample 1.550 0.922 0.519 0.312 0.218 0.053 -80°C, 21-day sample 2.190 1.966 1.224 0.784 0.504 0.057 37°C, 21-day sample 2.249 2.073 1.537 0.898 0.535 0.047 4°C, 21-day sample 2.031 1.927 1.138 0.745 0.542 0.039 -80°C, 21-day sample 2.132 1.985 1.441 0.796 0.649 0.038

[0145] 3. Stability test

[0146] The above antibodies were placed at 4℃ (refrigerator), -80℃ (refrigerator) and 37℃ (incubator) for 21 days, and samples taken at 7 days, 14 days and 21 days were observed for state, and the 21-day samples were tested for activity. The results showed that under the three test conditions, the antibodies were placed for 21 days, and no obvious change in protein state was observed, and the activity did not show a downward trend with the increase of test temperature, indicating that the above antibodies were stable. Table 5 below shows the OD results of enzyme immunoactivity detection of antibody Anti-AMH 15G10Rmb3 tested for 21 days.

[0147] Table 5 Stability data

[0148] 37°C, 21-day sample 23.438 11.719 0.000 4°C, 21-day sample 1.531 0.825 0.021 -80°C, 21-day sample 1.544 0.834 0.017 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C, 21-day sample 37°C, 21-day sample 4°C, 21-day sample -80°C 1.537 0.876 0.033

[0149] The preferred embodiments of the present application have been described above with the purpose of enabling not only the best modes contemplated by the inventors of carrying out the application, but also as examples of specific embodiments of the application. Numerous modifications and variations are possible in light of the above teachings. It is therefore to be understood that within the scope of the present application, the application can be practiced otherwise than as specifically described. Any reference to claimed subject matter presents a properly

[0150] Some of the amino acid sequences referred to in this application are shown below:

[0151]

[0152]

Claims

1. An antibody or antigen-binding fragment thereof that binds anti-Mullerian hormone, the antibody or antigen-binding fragment thereof comprising HCDR1, HCDR2, HCDR3, and LCDR1, LCDR2, LCDR3, characterized in that, The HCDR1, HCDR2, and HCDR3 are amino acid sequences identical to the HCDR1, HCDR2, and HCDR3 of the heavy chain variable region shown in SEQ ID NO: 17; the LCDR1, LCDR2, and LCDR3 are amino acid sequences identical to the LCDR1, LCDR2, and LCDR3 of the light chain variable region shown in any one of SEQ ID NOs: 19, 20, 21, and 22, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of the variable region are defined by any one of Kabat, Chothia, IMGT, AbM, or Contact.

2. An antibody or antigen-binding fragment thereof that binds anti-Müllerian hormone, characterized in that, The antibody or antigen-binding fragment thereof comprises the following complementarity determining regions: the amino acid sequence of HCDR1 is shown in SEQ ID NO: 1; the amino acid sequence of HCDR2 is shown in SEQ ID NO: 2; the amino acid sequence of HCDR3 is shown in SEQ ID NO: 3; the amino acid sequence of LCDR1 is shown in SEQ ID NO: 4 or 30; the amino acid sequence of LCDR2 is shown in SEQ ID NO: 5; the amino acid sequence of LCDR3 is shown in SEQ ID NO: 6 or 31.

3. The antibody or antigen-binding fragment thereof of claim 1 or 2, wherein, The antibody or antigen-binding fragment thereof further comprises HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4.

4. The antibody or antigen-binding fragment thereof according to claim 3, wherein, the amino acid sequence of HFR1 is SEQ ID NO: 7 or an amino acid sequence having at least 80% identity thereto; the amino acid sequence of HFR2 is SEQ ID NO: 8 or an amino acid sequence having at least 80% identity thereto; the amino acid sequence of HFR3 is SEQ ID NO: 9 or an amino acid sequence having at least 80% identity thereto; the amino acid sequence of HFR4 is SEQ ID NO: 10 or an amino acid sequence having at least 80% identity thereto; the amino acid sequence of LFR1 is SEQ ID NO: 11 or an amino acid sequence having at least 80% identity thereto; the amino acid sequence of LFR2 is SEQ ID NO: 12 or an amino acid sequence having at least 80% identity thereto; the amino acid sequence of LFR3 is SEQ ID NO: 13 or an amino acid sequence having at least 80% identity thereto; the amino acid sequence of LFR4 is SEQ ID NO: 14 or an amino acid sequence having at least 80% identity thereto.

5. The antibody or antigen-binding fragment thereof of any one of claims 1 to 2, 4, wherein, The antibody or antigen-binding fragment thereof binds to Anti-Mullerian hormone with a KD < 7.21 x 10 -7 M binds to Anti-Mullerian hormone with high affinity.

6. An antibody or antigen-binding fragment thereof that binds anti-Müllerian hormone, comprising a heavy chain variable region and a light chain variable region, characterized in that, The heavy chain variable region and the light chain variable region are selected from any one of the following combinations: 。 7. The antibody or antigen-binding fragment thereof of any one of claims 1 to 2, 4, 6, wherein, The antibody or antigen-binding fragment thereof further comprises a constant region.

8. The antibody or antigen-binding fragment thereof of claim 7, wherein, The constant region comprises a heavy chain constant region and / or a light chain constant region.

9. The antibody or antigen-binding fragment thereof of claim 8, wherein, The heavy chain constant region is selected from the heavy chain constant region of IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, or IgD; and the light chain constant region is selected from the kappa type or lambda type light chain constant region.

10. The antibody or antigen-binding fragment thereof of claim 7, wherein, The constant region is of bovine, equine, porcine, ovine, caprine, rat, mouse, dog, cat, rabbit, donkey, deer, mink, chicken, duck, goose, or human origin.

11. The antibody or antigen-binding fragment thereof of claim 7, wherein, The constant region is of mouse origin.

12. The antibody or antigen-binding fragment thereof of claim 8, wherein, The sequence of the heavy chain constant region is set forth in SEQ ID NO: 15 or has at least 80% identity thereto; the sequence of the light chain constant region is set forth in SEQ ID NO: 16 or has at least 80% identity thereto.

13. The antibody or antigen-binding fragment thereof of any one of claims 1 to 2, 4, 6, wherein, The antigen binding fragment is selected from any one of F(ab')2, Fab', Fab, Fv, and scFv of the antibody.

14. An antibody that binds anti-Müllerian hormone, comprising a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 1 and the light chain comprises the amino acid sequence of SEQ ID NO:

2. The amino acid sequence of the heavy chain and the amino acid sequence of the light chain are selected from any one of the following combinations: 。 15. An antibody conjugate, characterized in that, The antibody conjugate comprises the antibody or antigen binding fragment thereof of any one of claims 1 to 13 or the antibody of claim 14 and a label conjugated to the antibody or antigen binding fragment thereof.

16. The antibody conjugate of claim 15, wherein, The label is selected from biotin, a fluorescent dye, an enzyme, a radioisotope, a chemiluminescent reagent, and a nanoparticle-based label.

17. An antibody conjugate, characterized in that, The antibody conjugate comprises the antibody or antigen binding fragment thereof of any one of claims 1 to 13 or the antibody of claim 14 and a solid support conjugated to the antibody or antigen binding fragment thereof.

18. The antibody conjugate of claim 17, wherein, The solid support is selected from a microsphere, a plate, and a membrane.

19. A reagent or kit characterized in that, The reagent or kit comprises the antibody or antigen binding fragment thereof of any one of claims 1 to 13 or the antibody of claim 14 or the antibody conjugate of any one of claims 15 to 18.

20. Use of an antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, an antibody according to claim 14, or an antibody conjugate according to any one of claims 15 to 18 for the manufacture of a reagent or kit for the detection of anti-Mullerian hormone, characterized in that, Comprising: a) contacting the antibody or antigen binding fragment thereof of any one of claims 1 to 13, the antibody of claim 14, or the antibody conjugate of any one of claims 15 to 18 with anti-Mullerian hormone in a sample to be tested under conditions sufficient for an antibody / antigen binding reaction to occur to form an immunocomplex; and b) detecting the presence of the immunocomplex, the presence of the complex indicating the presence of the antigen in the test sample.

21. Use according to claim 20, characterized in that, The immunocomplex further comprises a second antibody that binds to the antibody or antigen binding fragment thereof.

22. The use according to claim 20, characterized in that, The immunocomplex further comprises a second antibody that binds to anti-Mullerian hormone.

23. A nucleic acid, comprising: which encodes the antibody or antigen binding fragment thereof of any one of claims 1 to 13 or the antibody of claim 14.

24. A vector, comprising: which contains the nucleic acid of claim 23.

25. A cell, comprising: which contains the nucleic acid of claim 23 or the vector of claim 24.

26. A method of preparing an antibody or antigen binding fragment thereof according to any one of claims 1 to 13 or an antibody according to claim 14, characterized in that, which comprises: culturing the cell of claim 25.

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