Anti-follicle-stimulating hormone antibody and application thereof

Abstract

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

Description

Technical Field

[0001] The present invention relates to the technical field of antibodies, and in particular to an anti-follicle stimulating hormone antibody and an application thereof. Background Art

[0002] Follicle-stimulating hormone (FSH) is a glycosylated protein hormone synthesized and secreted by the pituitary gland. It is a crucial hormone in both sexes, regulating a range of physiological processes related to development, growth, puberty, sexual maturation, and reproduction. In men, it acts on the seminiferous tubules of the testes to promote spermatogenesis and on the Sertoli cells to secrete estradiol. In women, it promotes the proliferation and differentiation of granulosa cells in the ovarian follicles, promoting ovarian growth. Specifically, when estrogen levels decrease, FSH is released from the pituitary gland, signaling the onset of menstruation or menopause. In the early stages of menopause, hormonal imbalances regulating the menstrual cycle occur. As women age beyond their reproductive years, ovarian estrogen production gradually decreases, leading to elevated FSH levels. FSH normally regulates egg growth and development. With the end of menstruation, FSH secretion ceases and returns to normal. As estrogen production decreases with age, follicle-stimulating hormone (FSH) production increases. These hormones gradually change over time, culminating in a complete cessation of menstrual cycles, a condition known as menopause. Two to ten years after the last menstrual period, ovarian function gradually deteriorates. The early stages before menopause are called perimenopause, during which FSH levels may rise and then slowly return to normal, leading to menstrual irregularities or menopause. Therefore, FSH and LH play a crucial role in reproductive processes, and regular testing should be performed to help determine FSH levels in the body, particularly during a woman's menstrual cycle.

[0003] At present, the detection method of FSH mainly contains colloidal gold immunochromatography (Immunochromatography Assay), which is a kind of immunological detection method based on antigen-antibody specific reaction.Its basic principle is: utilize colloidal gold to mark a kind of antigen or antibody, corresponding pairing antigen or antibody are coated on nitrocellulose membrane, when detecting sample, colloidal gold label combines with the part in sample to form complex, and then moves upward by chromatography, combines with coated antigen or antibody and condenses color development, thereby realizes the judgment to sample testing result.Other detects FSH the same with colloidal gold immunochromatography immunoassay, all need the antibody for FSH.Therefore, those skilled in the art have strong demand for the anti-FSH antibody with good performance. Summary of the Invention

[0004] The present application provides an anti-follicle stimulating hormone antibody, which provides an important source of raw materials for the detection of follicle stimulating hormone and has good activity or affinity.

[0005] In order to achieve the above object, according to one aspect of the present invention, an anti-follicle stimulating hormone antibody is provided, wherein the antibody comprises the following complementary determining regions:

[0006] (1) an amino acid sequence of HCDR1 as set forth in SEQ ID NO: 1, HCDR2 as set forth in SEQ ID NO: 2, HCDR3 as set forth in SEQ ID NO: 3 or SEQ ID NO: 17; and an amino acid sequence of LCDR1 as set forth in SEQ ID NO: 4 or SEQ ID NO: 18, LCDR2 as set forth in SEQ ID NO: 5, or LCDR3 as set forth in SEQ ID NO: 6; or

[0007] (2) The amino acid sequence of HCDR1 is as shown in SEQ ID NO:35, the amino acid sequence of HCDR2 is as shown in SEQ ID NO:36, the amino acid sequence of HCDR3 is as shown in SEQ ID NO:37 or SEQ ID NO:51; and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are as shown in SEQ ID NO:38, SEQ ID NO:39 and SEQ ID NO:40, respectively.

[0008] To achieve the above-mentioned object, according to a second aspect of the present invention, there is provided an anti-follicle-stimulating hormone antibody, 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:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, and SEQ ID NO:57; and the amino acid sequence of the light chain variable region is shown in any one of SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:64, and SEQ ID NO:65.

[0009] To achieve the above object, according to the third aspect of the present invention, an anti-follicle stimulating hormone antibody is provided, wherein the antibody comprises the three heavy chain complementary determining regions of any group of heavy chain variable regions and the three light chain complementary determining regions of the light chain variable region described in the second aspect.

[0010] To achieve the above-mentioned object, according to the fourth aspect of the present invention, there is provided an anti-follicle-stimulating hormone antibody, comprising a heavy chain and / or a light chain, wherein the amino acid sequence of the aforementioned heavy chain is shown in any one of SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, and SEQ ID NO:62; and the amino acid sequence of the aforementioned light chain is shown in any one of SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:66, and SEQ ID NO:67.

[0011] In order to achieve the above object, according to the fifth aspect of the present invention, an antibody conjugate is provided, wherein the antibody conjugate comprises the above antibody.

[0012] In order to achieve the above object, according to the sixth aspect of the present invention, a reagent or kit is provided, wherein the reagent or kit comprises the above antibody or the above antibody conjugate.

[0013] In order to achieve the above-mentioned object, according to the seventh aspect of the present invention, there is provided a use of the above-mentioned antibody or antibody conjugate in the preparation of a product for detecting follicle-stimulating hormone.

[0014] In order to achieve the above object, the present invention also provides a nucleic acid, a vector, a cell and a method for preparing the above antibody. DETAILED DESCRIPTION

[0015] In a first aspect, an embodiment of the present invention provides an anti-follicle stimulating hormone antibody, wherein the antibody comprises the following complementary determining regions:

[0016] (1) an amino acid sequence of HCDR1 as set forth in SEQ ID NO: 1, HCDR2 as set forth in SEQ ID NO: 2, HCDR3 as set forth in SEQ ID NO: 3 or SEQ ID NO: 17; and an amino acid sequence of LCDR1 as set forth in SEQ ID NO: 4 or SEQ ID NO: 18, LCDR2 as set forth in SEQ ID NO: 5, or LCDR3 as set forth in SEQ ID NO: 6; or

[0017] (2) The amino acid sequence of HCDR1 is as shown in SEQ ID NO:35, the amino acid sequence of HCDR2 is as shown in SEQ ID NO:36, the amino acid sequence of HCDR3 is as shown in SEQ ID NO:37 or SEQ ID NO:51; and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are as shown in SEQ ID NO:38, SEQ ID NO:39 and SEQ ID NO:40, respectively.

[0018] In the present invention, the term "antibody" is used in the broadest sense and may include full-length monoclonal antibodies, bispecific antibodies, multispecific antibodies, chimeric antibodies or antigen-binding fragments, so long as they exhibit the desired biological activity.

[0019] Antigen-binding fragments typically have the same binding specificity as the antibody from which they are derived. Those skilled in the art will readily appreciate, based on the disclosure herein, that such antigen-binding fragments can be obtained by methods such as enzymatic digestion (including pepsin or papain) and / or by chemical reduction to cleave disulfide bonds. Based on the structure of the intact antibody disclosed herein, those skilled in the art can readily obtain such antigen-binding fragments.

[0020] Antigen-binding fragments can also be synthesized by recombinant genetic techniques also known to those skilled in the art or by, for example, an automated peptide synthesizer, such as those sold by Applied BioSystems and the like.

[0021] As used herein, the terms "complementarity determining region," "CDR," or "CDRs" refer to the hypervariable regions of the heavy and light chains of immunoglobulins, and include one or more, or even all, of the amino acid residues that contribute substantially to the binding of an antibody or antigen-binding fragment to its recognized antigen or epitope. In specific embodiments of the present invention, CDRs refer to the hypervariable regions of the heavy and light chains of antibodies.

[0022] In the present invention, the heavy chain complementarity determining region is represented by HCDR, which includes HCDR1, HCDR2 and HCDR3; the light chain complementarity determining region is represented by LCDR, which includes LCDR1, LCDR2 and LCDR3.

[0023] Methods for defining CDRs are well known in the art, including 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 by Kabat et al., U.S. Patent No. 200200201154, "Sequence of Proteins of Immunological Interest" (1983). The "Chothia definition" refers to the definition system described by Chothia et al., J Mol Biol 196: 901-917 (1987). Other CDR definition methods may not strictly follow one of the above schemes, but will still overlap with at least a portion of the CDR region defined by Kabat, although they may be shortened or lengthened based on predictions or experimental results of specific residues or residue groups. Exemplary defined CDRs are listed in Table 1 below, and the definitions in different literature vary slightly. Given the variable region amino acid sequence of an antibody, a person skilled in the art can routinely determine which residues comprise a specific CDR. It should be noted that CDRs defined by other methods other than those in Table 1 also fall within the scope of protection of the present disclosure.

[0024] Table 1: CDR Definition 1

[0025] CDR Kabat AbM2 IMGT Chothia HCDR1 <![CDATA[H31~H35 3 ]]> <![CDATA[H26~H35 3 ]]> <![CDATA[H26~H33..5 5 ]]> <![CDATA[H26~H32..34 4 ]]> 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 LCDR3 L89 - L97 L89 - L97 L89 - L97 L89 - L97

[0026] 1 The numbering of all CDR definitions in Table 1 is based on the Kabat numbering system (see below), with amino acid numbers on the heavy chain represented by "H+number" and amino acid numbers on the light chain represented by "L+number." One of ordinary skill in the art can unambiguously assign this Kabat numbering system to any variable region sequence without relying on any experimental data other than the sequence itself. As used herein, "Kabat numbering" refers to the numbering system described in Kabat et al., U.S. Pat. of Health and Human Services, "Sequence of Proteins of Immunological Interest" (1983).

[0027] 2 "AbM" as used in Table 1 with a lowercase "b" refers to CDRs defined by Oxford Molecular's "AbM" antibody modeling software.

[0028] 3If both H35A and H35B are absent, CDR-H1 ends at position 35; if only H35A is present, CDR-H1 ends at position 35A; if both H35A and H35B are present, CDR-H1 ends at position 35B.

[0029] 4 If both H35A and H35B are absent, CDR-H1 ends at position 32; if only H35A is present, CDR-H1 ends at position 33; if both H35A and H35B are present, CDR-H1 ends at position 34.

[0030] 5 If both H35A and H35B are absent, CDR-H1 ends at position 33; if only H35A is present, CDR-H1 ends at position 34; if both H35A and H35B are present, CDR-H1 ends at position 35.

[0031] According to an embodiment of the present invention, the above-mentioned HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 or LCDR3 is defined by any one of the Kabat, Chothia, IMGT, AbM or Contact systems or a combination of multiple systems.

[0032] In some optional embodiments of the present invention, the above-mentioned HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the Kabat system.

[0033] In some optional embodiments of the present invention, the above-mentioned HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the Chothia system.

[0034] In some optional embodiments of the present invention, the above-mentioned HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the IMGT system.

[0035] In some optional embodiments of the present invention, the above-mentioned HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by the AbM system.

[0036] In some optional embodiments of the present invention, the above-mentioned HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by a Contact system.

[0037] In some optional embodiments of the present invention, the above-mentioned HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 are defined by a combination of Kabat, Chothia, IMGT, AbM or Contact systems.

[0038] In a second aspect, an embodiment of the present invention provides an anti-follicle stimulating hormone antibody, wherein the antibody comprises the following complementary determining regions:

[0039] (1) an amino acid sequence of HCDR1 as set forth in SEQ ID NO: 1, HCDR2 as set forth in SEQ ID NO: 2, HCDR3 as set forth in SEQ ID NO: 3 or SEQ ID NO: 17; and an amino acid sequence of LCDR1 as set forth in SEQ ID NO: 4 or SEQ ID NO: 18, LCDR2 as set forth in SEQ ID NO: 5, or LCDR3 as set forth in SEQ ID NO: 6; or

[0040] (2) The amino acid sequence of HCDR1 is as shown in SEQ ID NO:35, the amino acid sequence of HCDR2 is as shown in SEQ ID NO:36, the amino acid sequence of HCDR3 is as shown in SEQ ID NO:37 or SEQ ID NO:51; and the amino acid sequences of LCDR1, LCDR2 and LCDR3 are as shown in SEQ ID NO:38, SEQ ID NO:39 and SEQ ID NO:40, respectively.

[0041] According to an embodiment of the present invention, the above HCDRs and LCDRs are defined by the Kabat system.

[0042] In the present invention, the "framework region" or "FR" region includes the heavy chain framework region and the light chain framework region, and refers to the region of the antibody heavy chain variable region and the light chain variable region excluding CDR; wherein the heavy chain framework region can be further subdivided into adjacent regions separated by CDR, including HFR1, HFR2, HFR3 and HFR4 framework regions; the light chain framework region can be further subdivided into adjacent regions separated by CDR, including LFR1, LFR2, LFR3 and LFR4 framework regions.

[0043] In the present invention, the heavy chain variable region is obtained by arranging and connecting the following numbered CDRs and FRs in the following combinations: HFR1-HCDR1-HFR2-HCDR2-HFR3-HCDR3-HFR4; the light chain variable region is obtained by arranging and connecting the following numbered CDRs and FRs in the following combinations: LFR1-LCDR1-LFR2-LCDR2-LFR3-LCDR3-LFR4.

[0044] In an optional embodiment, the antibody of the first aspect further has HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3 and LFR4.

[0045] In an optional embodiment, the above antibody further comprises the following framework regions:

[0046] (1) HFR1, HFR2, HFR3, and HFR4 whose amino acid sequences are sequentially as shown in SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10; and LFR1, LFR2, LFR3, and LFR4 whose amino acid sequences are sequentially shown in SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, and SEQ ID NO:14; or amino acid sequences that are at least 80% identical to the foregoing framework region sequences; or

[0047] (2) HFR1, HFR2, HFR3, and HFR4 whose amino acid sequences are shown in SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, and SEQ ID NO:44, respectively; and LFR1, LFR2, LFR3, and LFR4 whose amino acid sequences are shown in SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, and SEQ ID NO:48, respectively; or amino acid sequences that are at least 80% identical to the aforementioned framework region sequences.

[0048] It should be noted that:

[0049] The framework region sequence of the antibody corresponds to the complementary determining region sequence described in the first aspect according to the numerical sequence. For example, if the framework region sequence of the antibody is shown as the amino acid sequence in sequence number (1), the corresponding framework region sequence is also shown as sequence number (1).

[0050] In other embodiments, the amino acid sequences of the framework regions of the anti-follicle stimulating hormone antibodies provided herein may be at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the corresponding framework regions (SEQ ID NO: 7, 8, 9, 10, 11, 12, 13 or 14).

[0051] In an alternative embodiment, the above antibody has a KD of <4.74×10 -9 Affinity of M for follicle-stimulating hormone.

[0052] In an optional embodiment, the above antibody has a KD≤10-8 M, KD≤10 -9 M, KD≤10 -10 M, KD≤10 -11 M or KD≤10 -12 M binds follicle-stimulating hormone with high affinity.

[0053] In an optional embodiment, the above antibody has a KD≤2.99×10 -9 M binds follicle-stimulating hormone with high affinity.

[0054] There are many methods for determining antibody affinity (KD). Based on the detection principle, they can be divided into thermodynamic detection methods, kinetic detection methods, and dynamic equilibrium detection methods. Among them, thermodynamic detection methods such as isothermal titration calorimetry (ITC) are common; kinetic detection methods such as surface plasmon resonance (SPR) and biofilm interferometry (BLI) are common; and dynamic equilibrium detection methods such as enzyme-linked immunosorbent assay (ELISA) are common.

[0055] In alternative embodiments, KD is determined using a kinetic assay; optionally, surface plasmon resonance, for example, by using a kinetic assay such as System of biosensor systems.

[0056] In a second aspect, an embodiment of the present invention provides an anti-follicle-stimulating hormone antibody, 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: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, and SEQ ID NO: 57, and the amino acid sequence of the light chain variable region is shown in any one of SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 64, and SEQ ID NO: 65.

[0057] In an optional embodiment, the heavy chain variable region and light chain variable region described in the second aspect are selected from any one of the following combinations:

[0058]

[0059]

[0060] In a third aspect, an embodiment of the present invention provides an anti-follicle stimulating hormone antibody, wherein the antibody comprises the three heavy chain complementary determining regions of any one group of heavy chain variable regions and the three light chain complementary determining regions of the light chain variable region in the second aspect.

[0061] In an optional embodiment, the complementarity determining regions described in the first or third aspect are defined by any one of the Kabat, Chothia, IMGT, AbM or Contact systems or a combination of multiple systems.

[0062] In an optional embodiment, the antibody described in the first, second or third aspect further comprises a constant region.

[0063] In an alternative embodiment, the above-mentioned constant region includes a heavy chain constant region and / or a light chain constant region.

[0064] In an optional embodiment, the heavy chain constant region is selected from any one of the heavy chain constant regions of IgG, IgA, IgM, IgE, and IgD, or a combination of multiple constant region segments.

[0065] In an optional embodiment, the heavy chain constant region includes CH1 of IgG, the hinge region of IgG, CH2 of IgM, CH3 of IgM and / or CH4 of IgM.

[0066] In an optional embodiment, the above-mentioned IgG is selected from IgG1, IgG2, IgG3 or IgG4.

[0067] In an alternative embodiment, the above-mentioned light chain constant region is selected from a kappa-type or lambda-type light chain constant region.

[0068] In an optional embodiment, the species origin of the above-mentioned constant region is cattle, horse, dairy cow, pig, sheep, rat, mouse, dog, camel, cat, rabbit, donkey, deer, mink, chicken, duck, goose, turkey, fighting cock or human.

[0069] In an optional embodiment, the species origin of the above-mentioned constant region is mouse.

[0070] In this article, the division of variable and constant region sequences refers to the IMGT division method, see Lefranc, and Martinez-Jean C. and Bosc N. or Ehrenmann,Patrice Duroux,Chantal Ginestoux,Gene table:housemouse(Mus musculus)IGHC,IMGT Repertoire. the internationalImMunoGenetics information http: / / www.imgt.org .Created:16 / 03 / 2011.Version:17 / 01 / 2020.or Ehrenmann,Patrice Duroux,Chantal Ginestoux,Gene table:house mouse(Mus musculus)IGLC,IMGT Repertoire. theinternational ImMunoGenetics information http: / / www.imgt.org .Created: 16 / 03 / 2011.Version: 17 / 01 / 2020.. Variable regions divided by different methods may differ from the variable region C-terminus or constant region N-terminus divided by IMGT. Variable regions or constant regions divided by other methods known in the art are also within the scope of protection of the present invention.

[0071] In an alternative embodiment, the above antibody comprises the following constant region:

[0072] (1) an amino acid sequence such as CH of SEQ ID NO: 15; and an amino acid sequence such as CL of SEQ ID NO: 16; or an amino acid sequence having at least 80% identity with each of the aforementioned constant regions; or

[0073] (2) an amino acid sequence such as CH of SEQ ID NO: 49; and an amino acid sequence such as CL of SEQ ID NO: 50; or an amino acid sequence having at least 80% identity with each of the aforementioned constant regions.

[0074] It should be noted that:

[0075] The amino acid sequence of the constant region of the antibody corresponds one-to-one with the complementary determining region and framework region described in the first aspect according to the numerical sequence. For example, if the amino acid sequence of the constant region of the antibody is as shown in the amino acid sequence in (1), then the corresponding complementary determining region and framework region are also as shown in the amino acid sequence shown in (1).

[0076] In other embodiments, the constant region sequence can be at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the constant region (SEQ ID NO: 15 or 16).

[0077] In an optional embodiment, the above-mentioned antibody includes any one of F(ab)2, F(ab')2, Fab', Fab, Fv and scFv.

[0078] In a fourth aspect, the present invention provides an anti-follicle-stimulating hormone antibody, 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:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, and SEQ ID NO:62, and the amino acid sequence of the light chain is shown in any one of SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:66, and SEQ ID NO:67.

[0079] In an optional embodiment, the antibody of the first, second, third or fourth aspect comprises any combination of the following heavy chains and light chains:

[0080]

[0081]

[0082] In a fifth aspect, the present invention provides an antibody conjugate, wherein the antibody conjugate comprises the above-mentioned antibody.

[0083] In an optional embodiment, the antibody conjugate comprises biotin or a biotin derivative.

[0084] In an optional embodiment, the above-mentioned antibody conjugate further comprises a marker or a purification tag.

[0085] In an optional embodiment, the above-mentioned marker refers to a class of substances with properties that can be directly observed by the naked eye or detected or detected by an instrument, such as luminescence, color development, radioactivity, etc., through which qualitative or quantitative detection of the corresponding target can be achieved.

[0086] In an optional embodiment, the above-mentioned labels include but are not limited to fluorescent dyes, enzymes, radioactive isotopes, chemiluminescent reagents and nanoparticle labels.

[0087] In actual use, those skilled in the art can select a suitable marker according to the detection conditions or actual needs. No matter which marker is used, it falls within the scope of protection of the present invention.

[0088] In an optional embodiment, the fluorescent dyes include but are not limited to fluorescein dyes and their derivatives (for example, including but not limited to fluorescein isothiocyanate (FITC), hydroxyfluorescein (FAM), tetrachlorofluorescein (TET), etc. or their analogs), rhodamine dyes and their derivatives (for example, including but not limited to red rhodamine (RBITC), tetramethylrhodamine (TAMRA), rhodamine B (TRITC), etc. or their analogs), Cy series dyes and their derivatives (for example, including but not limited to Cy2, Cy3, Cy3B, Cy3.5, C y5, Cy5.5, Cy3, etc. or their analogs), Alexa series dyes and their derivatives (for example, including but not limited to AlexaFluor350, 405, 430, 488, 532, 546, 555, 568, 594, 610, 33, 647, 680, 700, 750, etc. or their analogs) and protein dyes and their derivatives (for example, including but not limited to phycoerythrin (PE), phycocyanin (PC), allophycocyanin (APC), peridinin-chlorophyll protein (preCP), etc.).

[0089] In an optional embodiment, the enzymes include but are not limited to horseradish peroxidase, alkaline phosphatase, β-galactosidase, glucose oxidase, carbonic anhydrase, acetylcholinesterase and 6-phosphate glucose deoxidase.

[0090] In an optional embodiment, the above-mentioned radioactive isotopes include but are 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.

[0091] In an optional embodiment, the above-mentioned chemiluminescent reagents include but are not limited to luminol and its derivatives, lucigenin, crustacean fluorescein and its derivatives, ruthenium bipyridine and its derivatives, acridinium esters and their derivatives, dioxetanes and their derivatives, lophanes and their derivatives, and peroxalates and their derivatives.

[0092] In an optional embodiment, the above-mentioned nanoparticle markers include but are not limited to nanoparticles, colloids, organic nanoparticles, magnetic nanoparticles, quantum dot nanoparticles and rare earth complex nanoparticles.

[0093] In an optional embodiment, the colloid includes but is not limited to colloidal metal, colloidal carbon, disperse dyes, dye-labeled microspheres and latex.

[0094] In an optional embodiment, the colloidal metal includes but is not limited to colloidal gold, colloidal silver and colloidal selenium.

[0095] In an optional embodiment, the colloidal metal is colloidal gold.

[0096] In an optional embodiment, the antibody conjugate further comprises a solid phase carrier coupled to the antibody.

[0097] In an optional embodiment, the solid phase carrier is selected from microspheres, plates and membranes.

[0098] In an optional embodiment, the solid phase carrier includes but is not limited to magnetic microspheres, plastic microspheres, plastic particles, microporous plates, glass, capillaries, nylon and nitrocellulose membranes.

[0099] In a sixth aspect, the present invention provides a reagent or a kit, wherein the reagent or the kit comprises the above-mentioned antibody or the above-mentioned antibody conjugate.

[0100] As previously mentioned, the antibodies described in some embodiments or examples of the present invention can effectively bind to follicle-stimulating hormone (FSH). Therefore, reagents or kits containing these FSH antibodies can effectively detect FSH qualitatively or quantitatively. The reagents or kits provided by the present invention can be used, for example, in immunoblotting, immunoprecipitation, and other assays that utilize the specific binding properties of FSH and its antibodies. As previously mentioned, the antibodies described in some embodiments or examples of the present invention have higher binding activity or affinity for FSH. Therefore, reagents or kits containing these antibodies have higher detection sensitivity or specificity.

[0101] In a seventh aspect, the present invention provides a method for detecting follicle-stimulating hormone, comprising: a) contacting the above-mentioned antibody, antibody conjugate, reagent or kit with follicle-stimulating hormone in a sample to be tested under conditions sufficient for an antibody / antigen binding reaction to occur, to form an immune complex; and b) detecting the presence of the immune complex, wherein the presence of the immune complex indicates the presence of the antigen in the test sample;

[0102] In an optional embodiment, the immune complex further comprises a second antibody, and the second antibody binds to the antibody.

[0103] In an optional embodiment, the immune complex further comprises a second antibody, and the second antibody binds to follicle-stimulating hormone.

[0104] In an eighth aspect, the present invention provides use of the above-mentioned anti-FSH antibodies and antibody conjugates in the preparation of products for detecting FSH.

[0105] It should be noted that the products of the present invention include but are not limited to reagents, test kits, test strips or reagent plates.

[0106] In a ninth aspect, the present invention provides a nucleic acid molecule encoding the above-mentioned antibody.

[0107] In a tenth aspect, the present invention provides a vector comprising the above-mentioned nucleic acid molecule.

[0108] In an eleventh aspect, the present invention provides a cell containing the above-mentioned vector.

[0109] In a twelfth aspect, the present invention provides a method for preparing an anti-follicle stimulating hormone antibody, comprising: culturing the cells as described above.

[0110] Based on the amino acid sequence of the anti-FSH antibody disclosed in the present invention, those skilled in the art can easily conceive of using genetic engineering technology or other technologies (chemical synthesis, recombinant expression) to prepare the anti-FSH antibody. For example, the antibody can be isolated and purified from the culture product of recombinant cells that can recombinantly express any of the above-mentioned antibodies. This is easy to achieve for those skilled in the art. Based on this, no matter what technology is used to prepare the anti-FSH antibody of the present invention, it falls within the scope of protection of the present invention.

[0111] To make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are described clearly and completely below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer are used. Where the manufacturer of the reagents or instruments is not specified, all are conventional products that can be purchased commercially.

[0112] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled 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. Unless otherwise indicated, the techniques employed or contemplated herein are standard methods. Materials, methods, and examples are illustrative and non-limiting only.

[0113] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of cell biology, molecular biology (including recombinant techniques), microbiology, biochemistry, and immunology, which are within the capabilities of a person skilled in the art. The technique is fully explained in the literature, for example, in Molecular Cloning: A Laboratory Manual, 2nd ed. (Sambrook et al., 1989); Oligonucleotide Synthesis (MJ Gait, ed., 1984); Animal Cell Culture (RI Freshney, ed., 1987); Methods in Enzymology (Academic Press, Inc.); Handbook of Experimental Immunology (DM Weir and CC Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (JM Miller and MP Calos, eds., 1987); Current Protocols in Molecular Biology (FM Ausubel et al., eds., 1987); and PCR: The Polymerase Chain Reaction. Reaction" (Mullis et al., eds., 1994); and Current Protocols in Immunology (JE Coligan et al., eds., 1991), each of which is expressly incorporated herein by reference.

[0114] The features and performance of the present invention are further described in detail below with reference to the embodiments.

[0115] Example 1 Antibody Discovery of Monoclonal Antibodies

[0116] 1. Animal immunization

[0117] FSH antigen (from Feipeng Bio) was mixed with equal volumes of Freund's complete adjuvant to produce an oily emulsion. This emulsion was injected subcutaneously into BALB / c mice at multiple sites at a dose of 0.2 ml per mouse. Fourteen days later, mice were immunized intraperitoneally with the same antigen and adjuvant. After the fourth injection, tail blood was collected for titer testing. Three days before fusion, mice were boosted intraperitoneally with the same dose of antigen mixed with an equal volume of 0.9% sodium chloride injection until the desired fusion titer was reached.

[0118] 2. Preparation of hybridoma cell lines

[0119] On the third day after the booster immunization, the spleens of the mice were removed under sterile conditions. Mouse tumor cells and immune spleen cells were mixed at a cell ratio of 1:10, fused, and cultured. On the sixth day of culture, the HT culture medium was changed twice. On the seventh day after fusion, the cell supernatant was collected for antibody detection to screen for hybridoma cell lines secreting specific antibodies. Two hybridoma cell lines were identified, designated Anti-FSH 9H7 and Anti-FSH 5C11.

[0120] Example 2 Preparation of monoclonal antibodies

[0121] In this example, restriction endonucleases and Prime Star DNA polymerase were purchased from Takara. MagExtractor-RNA extraction kit was purchased from TOYOBO. BD SMART TM The RACE cDNA Amplification Kit was purchased from Takara. The pMD-18T vector was also purchased from Takara. The plasmid extraction kit was purchased from Tiangen. Primer synthesis and gene sequencing were performed by Invitrogen.

[0122] 1. Antibody sequence acquisition

[0123] (1) Antibody gene preparation

[0124] mRNA was extracted from the hybridoma cell lines of Anti-FSH 9H7 and Anti-FSH 5C11. DNA products were obtained by RT-PCR. The products were subjected to A-addition reaction with rTaq DNA polymerase and inserted into the pMD-18T vector. The cells were transformed into DH5α competent cells. After colonies grew, the Heavy Chain and Light Chain gene clones were obtained. Four clones each were sent to a gene sequencing company for sequencing.

[0125] (2) Sequence analysis of antibody variable region genes

[0126] The gene sequences obtained by the above sequencing were placed in the Kabat antibody database for analysis, and VNTI11.5 software was used for analysis to confirm that the genes amplified by the heavy chain and light chain primer pairs were correct.

[0127] (3) Construction of recombinant antibody expression plasmid

[0128] pcDNA TM 3.4 The vector is a recombinant antibody eukaryotic expression vector constructed by PCR. Multiple cloning restriction sites such as HindIII, BamHI, and EcoRI have been introduced into the expression vector, and it is named pcDNA3.4A expression vector, hereinafter referred to as 3.4A expression vector. Based on the sequencing results of the antibody variable region gene in the above-mentioned pMD-18T, the VL and VH gene-specific primers of the antibody were designed, with HindIII and EcoRI restriction sites and protective bases at both ends, respectively. The light chain gene fragment and the heavy chain gene fragment were amplified by PCR.

[0129] The Heavy Chain and Light Chain gene fragments were digested with HindIII / EcoRI, and the 3.4A vector was digested with HindIII / EcoRI. After the fragments and vectors were purified and recovered, the Heavy Chain gene and Light Chain gene were respectively connected to the 3.4A expression vector to obtain the recombinant expression plasmids of Heavy Chain and Light Chain, respectively.

[0130] 2. Recombinant Antibody Production

[0131] Resuscitate HEK293 cells in advance and subculture them into 200 ml system to make the cell density reach 3-5×10 6 cells / ml cell density reaches the selected antibody concentration and cells, cell viability>95%; centrifuge and wash the cells, re-dissolve with culture medium, and adjust the cell density to 2.9×10 6 Cells were washed with 100 cells / ml and re-dissolved in culture medium, which was also used as a cell diluent. Plasmid DNA and transfection reagent diluents were prepared separately using culture medium. The transfection reagent diluent was added to the plasmid DNA diluent, mixed thoroughly, and allowed to stand at room temperature for 15 minutes. This mixture was slowly added to the cell diluent over 1 minute, mixed thoroughly, and samples were taken and counted. The viability of the cells after transfection was recorded and observed, and the cells were placed in a 35°C constant temperature incubator at 120 rpm and 8% CO2. After 13 days, the samples were collected by centrifugation. The supernatant was affinity purified using a protein A affinity chromatography column to obtain purified antibodies.

[0132] The obtained antibodies were named Anti-FSH 9H7Rmb1 and Anti-FSH 5C11Rmb1. Anti-FSH 9H7Rmb1 and Anti-FSH 5C11Rmb1 were mutated to obtain mutant antibodies. The sequences of the heavy chain (H) and light chain (L) of the above antibodies are shown in the following table:

[0133] Table 2: Antibody sequences

[0134] Antibody Name Heavy Chain Light Chain Anti - FSH 9H7RMb1 SEQ ID NO:24 SEQ ID NO:32 Anti - FSH 9H7RMb2 SEQ ID NO:28 SEQ ID NO:32 Anti - FSH 9H7RMb3 SEQ ID NO:25 SEQ ID NO:32 Anti - FSH 9H7RMb4 SEQ ID NO:27 SEQ ID NO:32 Anti - FSH 9H7RMb5 SEQ ID NO:26 SEQ ID NO:32 Anti - FSH 9H7RMb6 SEQ ID NO:24 SEQ ID NO:34 Anti - FSH 9H7RMb7 SEQ ID NO:24 SEQ ID NO:33 Anti - FSH 5C11RMb1 SEQ ID NO:58 SEQ ID NO:66 Anti - FSH 5C11RMb2 SEQ ID NO:59 SEQ ID NO:66 Anti - FSH 5C11RMb3 SEQ ID NO:62 SEQ ID NO:66 Anti - FSH 5C11RMb4 SEQ ID NO:63 SEQ ID NO:66 Anti - FSH 5C11RMb5 SEQ ID NO:58 SEQ ID NO:67 Anti - FSH 5C11RMb6 SEQ ID NO:61 SEQ ID NO:66 Anti - FSH 5C11RMb7 SEQ ID NO:60 SEQ ID NO:66 Anti - FSH 5C11RMb8 SEQ ID NO:63 SEQ ID NO:67

[0135] Example 2 Performance testing of antibodies

[0136] 1. Affinity Analysis

[0137] Purified antibodies were diluted in advance, and recombinant FSH antigen (from Feipeng Bio) was serially diluted. Binding and dissociation curves were measured on a Biacore 8K+ instrument using a CM5 chip pre-coupled with goat anti-mouse IgG. The instrument automatically fitted the affinity constant, association rate, and dissociation rate. (KD represents the equilibrium dissociation constant, also known as the affinity constant; ka represents the association rate; and kd represents the dissociation rate.)

[0138] Table 3: Affinity data

[0139] Sample Name KD(M) ka kd Control 4.74E-09 2.15E+04 1.02E-04 Anti - FSH 9H7RMb1 5.74E-10 5.85E+04 3.36E-05 Anti - FSH 9H7RMb2 4.17E-10 5.71E+04 2.38E-05 Anti - FSH 9H7RMb3 4.23E-10 5.81E+04 2.46E-05 Anti - FSH 9H7RMb4 4.27E-10 5.78E+04 2.47E-05 Anti - FSH 9H7RMb5 2.99E-09 1.98E+04 5.92E-05 Anti - FSH 9H7RMb6 6.01E-10 7.01E+04 4.21E-05 Anti - FSH 9H7RMb7 5.00E-10 3.52E+04 1.76E-05 Anti - FSH 5C11RMb1 4.55E-10 2.01E+05 9.15E-05 Anti-FSH 5C11RMb2 4.78E-10 1.88E+05 8.99E-05 Anti-FSH 5C11RMb3 5.05E-10 1.67E+05 8.43E-05 Anti-FSH 5C11RMb4 4.02E-10 2.66E+05 1.07E-04 Anti-FSH 5C11RMb5 6.87E-10 7.25E+05 4.98E-04 Anti-FSH 5C11RMb6 5.98E-10 6.69E+05 4.00E-04 Anti-FSH 5C11RMb7 1.06E-09 3.13E+05 3.31E-04 Anti-FSH 5C11RMb8 4.07E-10 1.01E+06 4.13E-04

[0140] 2. Activity Identification

[0141] (1) Dilute FSH recombinant antigen (from Feipeng Bio) to 3ug / ml in coating solution (main component: NaHCO3), add 100uL to each well, and incubate at 4℃ overnight.

[0142] (2) The next day, wash twice with washing solution (main components Na2HPO4 + NaCl) and pat dry; add blocking solution (20% BSA + 80% PBS), 120uL per well, incubate at 37℃ for 1 hour, and pat dry.

[0143] (3) Antibodies Anti-FSH 9H7RMb1 to Anti-FSH 9H7RMb7 were serially diluted from 71.825 ng / ml to 4.883 ng / ml in four steps; Antibodies Anti-FSH 5C11RMb1 to Anti-FSH 5C11RMb8 were serially diluted from 125 ng / ml to 7.81 ng / ml in four steps.

[0144] (4) Add 100 μL / well of the antibody diluted in step (3) and incubate at 37°C for 30 min. Wash 5 times with washing buffer and pat dry.

[0145] (5) Add goat anti-mouse IgG-HRP, 100 μL per well, 37°C, 30 min; wash 5 times with washing solution and pat dry.

[0146] (6) Add chromogenic solution A (50 μL / well), add chromogenic solution B (50 μL / well), incubate for 10 min; add stop solution (50 μL / well);

[0147] The OD value was read at 450 nm (reference 630 nm) on a microplate reader.

[0148] The experimental results are shown in Table 4 and Table 5 respectively.

[0149] Note: Solution A (main ingredients: citric acid + sodium acetate + acetanilide + urea peroxide); Solution B (main ingredients: citric acid + EDTA·2Na + TMB + concentrated HCl); Stop solution (EDTA·2Na + concentrated H2SO4)

[0150] Table 4: Activity data 1

[0151] Concentration (ng / ml) 78.125 39.063 19.531 9.766 4.883 0 Anti-FSH 9H7RMb1 1.65 1.179 0.821 0.475 0.274 0.035 Anti-FSH 9H7RMb2 1.639 1.178 0.826 0.484 0.269 0.031 Anti-FSH 9H7RMb3 1.643 1.175 0.834 0.477 0.277 0.038 Anti-FSH 9H7RMb4 1.645 1.181 0.831 0.483 0.271 0.027 Anti-FSH 9H7RMb5 2.127 1.568 0.903 0.491 0.281 0.071 Anti-FSH 9H7RMb6 1.896 1.189 0.659 0.374 0.292 0.033 Anti-FSH 9H7RMb7 1.831 1.271 0.908 0.439 0.204 0.01

[0152] Table 5: Activity data 1

[0153] Concentration (ng / ml) 125 62.5 31.25 15.63 7.81 0 Anti-FSH 5C11RMb1 1.909 1.336 0.792 0.365 0.174 0.022 Anti-FSH 5C11RMb2 1.808 1.367 0.732 0.394 0.156 0.013 Anti-FSH 5C11RMb3 1.982 1.378 0.758 0.357 0.132 0.012 Anti-FSH 5C11RMb4 1.825 1.301 0.775 0.405 0.174 0.019 Anti-FSH 5C11RMb5 1.877 1.269 0.727 0.444 0.228 0.028 Anti-FSH 5C11RMb6 1.883 1.373 0.718 0.351 0.234 0.037 Anti-FSH 5C11RMb7 1.879 1.368 0.723 0.455 0.239 0.023 Anti-FSH 5C11RMb8 1.865 1.245 0.885 0.435 0.151 0.013

[0154] Conclusion: Antibodies Anti-FSH 9H7RMb1~Anti-FSH 9H7RMb7 and Anti-FSH 5C11RMb1~Anti-FSH 5C11RMb8 have good activity.

[0155] 3. Stability assessment

[0156] The antibodies were placed at 4°C (refrigerator), -80°C (freezer), and 37°C (incubator) for 21 days. Samples were collected at 7, 14, and 21 days for status observation, and the 21-day sample was tested for activity. Tables 6 and 7 below show the OD results of enzyme immunoassay activity assays for Anti-FSH9H7RMb3 and Anti-FSH 5C11RMb7, respectively, over 21 days.

[0157] Table 6 Stability Data 1

[0158] Sample concentration (ng / ml) 39.063 19.531 0 4℃, 21-day samples 1.165 0.835 0.131 -80℃, 21-day sample 1.171 0.841 0.126 37℃, 21-day samples 1.163 0.834 0.135

[0159] Table 7 Stability Data 2

[0160]

[0161]

[0162] Conclusion: The results showed that no obvious changes in protein status were observed after 21 days of storage under the three test conditions, and the activity did not show a downward trend with the increase of the test temperature, indicating that the above antibodies are stable.

[0163] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

[0164] Some of the amino acid sequences involved in this application are shown in Table 8:

[0165]

[0166]

[0167]

[0168]

[0169]

[0170]

Claims

1. An anti-follicle stimulating hormone antibody, characterized in that The antibody comprises the following complementarity determining regions: (1) an amino acid sequence of HCDR1 as set forth in SEQ ID NO: 1, HCDR2 as set forth in SEQ ID NO: 2, HCDR3 as set forth in SEQ ID NO: 3 or SEQ ID NO: 17; and an amino acid sequence of LCDR1 as set forth in SEQ ID NO: 4 or SEQ ID NO: 18, LCDR2 as set forth in SEQ ID NO: 5, or LCDR3 as set forth in SEQ ID NO: 6; or (2) an amino acid sequence of HCDR1 as set forth in SEQ ID NO:35, HCDR2 as set forth in SEQ ID NO:36, HCDR3 as set forth in SEQ ID NO:37 or SEQ ID NO:51; and an amino acid sequence of LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NO:38, SEQ ID NO:39, and SEQ ID NO:40, respectively; Optionally, the antibody further comprises the following framework regions: (1) HFR1, HFR2, HFR3, and HFR4 whose amino acid sequences are sequentially as shown in SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10; and LFR1, LFR2, LFR3, and LFR4 whose amino acid sequences are sequentially shown in SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, and SEQ ID NO:14; or amino acid sequences that are at least 80% identical to said framework region sequences; or (2) HFR1, HFR2, HFR3, and HFR4 whose amino acid sequences are sequentially as shown in SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, and SEQ ID NO:44; and LFR1, LFR2, LFR3, and LFR4 whose amino acid sequences are sequentially shown in SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, and SEQ ID NO:48; or amino acid sequences that are at least 80% identical to the aforementioned framework region sequences; Optionally, the antibody has a KD of <4.74×10 -9 M binds follicle-stimulating hormone with high affinity.

2. An anti-follicle-stimulating hormone antibody comprising a heavy chain variable region and / or a light chain variable region, characterized in that: The amino acid sequence of the heavy chain variable region is shown in any one of SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, and SEQ ID NO: 57; the amino acid sequence of the light chain variable region is shown in any one of SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 64, and SEQ ID NO: 65; Optionally, the combination of the heavy chain variable region and the light chain variable region is selected from any one of the following combinations:

3. An anti-follicle stimulating hormone antibody, comprising three heavy chain complementarity determining regions of any one group of heavy chain variable regions and three light chain complementarity determining regions of a light chain variable region according to claim 2; Optionally, the complementarity determining regions of the variable regions are defined by any one of the Kabat, Chothia, IMGT, AbM or Contact systems or a combination of multiple systems; Optionally, the antibody further comprises a constant region; Optionally, the constant region includes a heavy chain constant region and / or a light chain constant region; Optionally, the heavy chain constant region is selected from any one of the heavy chain constant regions of IgG, IgA, IgM, IgE, and IgD, or a combination of multiple constant region segments; Optionally, the heavy chain constant region includes CH1 of IgG, hinge region of IgG, CH2 of IgM, CH3 of IgM and / or CH4 of IgM; Optionally, the species origin of the constant region is cow, horse, pig, sheep, goat, rat, mouse, dog, camel, cat, rabbit, donkey, deer, mink, chicken, duck, goose or human; Optionally, the species origin of the constant region is mouse; Optionally, the antibody comprises the following constant region: (1) an amino acid sequence such as CH of SEQ ID NO: 15; and an amino acid sequence such as CL of SEQ ID NO: 16; or an amino acid sequence having at least 80% identity with each of said constant regions; or (2) an amino acid sequence such as CH of SEQ ID NO: 49; and an amino acid sequence such as CL of SEQ ID NO: 50; or an amino acid sequence having at least 80% identity with each of said constant regions; Optionally, the antibody comprises any one of F(ab')2, Fab', Fab, Fv and scFv.

4. An anti-follicle stimulating hormone antibody comprising a heavy chain and / or a light chain, characterized in that: The amino acid sequence of the heavy chain is shown in any one of SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, and SEQ ID NO:62; the amino acid sequence of the light chain is shown in any one of SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:66, and SEQID NO:

67.

5. An antibody conjugate, characterized in that The antibody conjugate comprises the antibody according to any one of claims 1 to 4; Optionally, the antibody conjugate further comprises biotin or a biotin derivative conjugated to the antibody; Optionally, the antibody conjugate further comprises a marker or purification tag coupled to the antibody; Optionally, the label is selected from fluorescent dyes, enzymes, radioisotopes, chemiluminescent reagents and nanoparticle labels; Optionally, the antibody conjugate further comprises a solid phase carrier coupled to the antibody.

6. A reagent or kit, characterized in that The reagent or kit comprises the antibody according to any one of claims 1 to 4 or the antibody conjugate according to claim 5.

7. Use of the antibody according to any one of claims 1 to 4 or the antibody conjugate according to claim 5 in the preparation of a product for detecting follicle-stimulating hormone; Optionally, the use includes: a) contacting the antibody according to any one of claims 1 to 4, the antibody conjugate according to claim 5, or the reagent or kit according to claim 6 with follicle-stimulating hormone in a sample to be tested under conditions sufficient for an antibody / antigen binding reaction to form an immune complex; and b) detecting the presence of the immune complex, wherein the presence of the complex indicates the presence of the antigen in the test sample; Optionally, the immune complex further comprises a second antibody, which binds to the antibody; Optionally, the immune complex further comprises a second antibody, which binds to follicle-stimulating hormone.

8. A nucleic acid, a vector, a cell, or a method for producing the antibody of any one of claims 1 to 4, wherein the nucleic acid encodes the antibody of any one of claims 1 to 4; the vector comprises a nucleic acid encoding the antibody of any one of claims 1 to 4; the cell comprises the nucleic acid or vector; and the method comprises the cell.

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