Anti-HER-2 monoclonal antibody, composition and application thereof

Abstract

The invention relates to the field of biological detection, in particular to an anti-HER-2 monoclonal antibody, a composition and application thereof. The invention provides an anti-HER-2 monoclonal antibody, which comprises a monoclonal antibody 1 or a monoclonal antibody 2. The invention provides the HER-2 monoclonal antibody which is obtained by obtaining a hybridoma cell strain through a hybridoma technology, injecting the hybridoma cell strain into the abdominal cavity of a mouse to prepare ascites and purifying the ascites, the HER-2 monoclonal antibody can be prevented from being interfered by drugs, and the accuracy of immunodetection is effectively improved.

Description

Technical Field

[0001] This invention relates to the field of biological detection, and more particularly to anti-HER-2 monoclonal antibodies, compositions thereof, and their applications. Background Technology

[0002] Human epidermal growth factor receptor 2 (HER-2) is a member of the epidermal growth factor receptor (EGFR) family, a transmembrane tyrosine kinase receptor encoded by the erbB-2 / neu proto-oncogene. This gene is located on chromosome 17q21, and its encoded protein consists of 1255 amino acids with a molecular weight of 185 kDa, abbreviated as p185. Under normal physiological conditions, HER-2 precisely regulates key life activities such as cell growth, differentiation, and survival by forming heterodimers with other HER family members. However, when the HER-2 gene amplifies or its encoded protein is overexpressed, it leads to abnormal aggregation of HER-2 receptors on the cell membrane, triggering sustained and uncontrolled activation of downstream signaling pathways (such as the PI3K / Akt and Ras / MAPK pathways), thereby driving the malignant proliferation, invasion, and metastasis of tumor cells. This oncogenic mechanism has been confirmed in various solid tumors, with the most typical and in-depth studies found in breast cancer and gastric cancer.

[0003] Breast cancer (BC), one of the most common malignant tumors in women, poses a significant threat to women's health. Since the late 20th century, when the first monoclonal antibody targeting HER-2 was used for treatment, HER-2 targeted therapy has made significant progress. Currently, trastuzumab and pertuzumab are mainly used as treatments for HER-2 positive tumors. Pertuzumab is a humanized monoclonal antibody that binds to the HER-2 extracellular domain (ECD), inhibiting HER-2 heterodimer formation and its signal transduction activity, and reducing the activation of the second segment of the MAPK and Pl3K / Akt pathways, thereby inhibiting tumor cell survival, proliferation, and migration. However, these targeted antibody drugs can interfere with the detection of HER-2 protein in immunodiagnostics. Therefore, the development of HER-2 monoclonal antibodies against drug interference is of great significance for the diagnosis of breast cancer and other malignant tumors. Summary of the Invention

[0004] In view of this, the present invention provides an anti-HER-2 monoclonal antibody, a composition thereof, and its application. The present invention provides a HER-2 monoclonal antibody obtained by obtaining a hybridoma cell line through hybridoma technology, injecting the hybridoma cell line into the peritoneal cavity of mice to prepare ascites fluid, and then purifying the purified fluid. This antibody is immune to drug interference and effectively improves the accuracy of immune detection.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0006] The present invention provides a monoclonal antibody against HER-2, comprising: monoclonal antibody 1 or monoclonal antibody 2;

[0007] In the monoclonal antibody 1:

[0008] (1) The amino acid sequences of the three CDR regions of its heavy chain have the amino acid sequences shown in SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:11, respectively; and

[0009] (2) The amino acid sequences of the three CDR regions of its light chain have the amino acid sequences shown in SEQ ID NO:12, SEQ ID NO:13 and SEQ ID NO:14, respectively; or

[0010] (3) An amino acid sequence obtained by substituting, deleting, or adding one or more amino acids to the amino acid sequence described in (1) or (2), and which has the same or similar function as the amino acid sequence shown in (1) or (2); or

[0011] (4) An amino acid sequence that is at least 80% homologous to the sequence described in (1), (2) or (3); and / or

[0012] In the monoclonal antibody 2:

[0013] (5) The amino acid sequences of the three CDR regions of its heavy chain have the amino acid sequences shown in SEQ ID NO:15, SEQ ID NO:16 and SEQ ID NO:17, respectively; and

[0014] (6) The amino acid sequences of the three CDR regions of its light chain have the amino acid sequences shown in SEQ ID NO:18, SEQ ID NO:19 and SEQ ID NO:20, respectively; or

[0015] Amino acid sequences obtained by substituting, deleting, or adding one or more amino acids to the amino acid sequences described in (7), (5), or (6), and which have the same or similar function as the amino acid sequences shown in (5) or (6); or

[0016] (8) An amino acid sequence that is at least 80% homologous to the sequence described in (5), (6) or (7).

[0017] In some embodiments of the present invention, the above-mentioned monoclonal antibodies,

[0018] In the monoclonal antibody 1:

[0019] (9) Its heavy chain variable region has the amino acid sequence shown in SEQ ID NO:1; and

[0020] (10) Its light chain variable region has an amino acid sequence as shown in SEQ ID NO:2; or

[0021] Amino acid sequences obtained by substituting, deleting, or adding one or more amino acids to the amino acid sequences described in (11), (9), or (10), and which have the same or similar function as the amino acid sequences shown in (9) or (10); or

[0022] (12) An amino acid sequence that is at least 80% homologous to the sequence described in (9), (10) or (11); and / or

[0023] In the monoclonal antibody 2:

[0024] (13) Its heavy chain variable region has the amino acid sequence shown in SEQ ID NO:3; and

[0025] (14) Its light chain variable region has an amino acid sequence as shown in SEQ ID NO:4; or

[0026] Amino acid sequences obtained by substituting, deleting, or adding one or more amino acids to the amino acid sequences described in (15), (13), or (14), and which have the same or similar function as the amino acid sequences shown in (13) or (14); or

[0027] (16) An amino acid sequence that is at least 80% homologous to the sequences described in (13), (14) or (15).

[0028] The present invention also provides antibody conjugates comprising: the above-described monoclonal antibody and a detectable label.

[0029] The present invention also provides a composition of monoclonal antibodies, comprising: the monoclonal antibody 1 and the monoclonal antibody 2 of the above-described monoclonal antibody and / or the antibody conjugate described above.

[0030] The present invention also provides nucleic acid molecules encoding the above-described monoclonal antibodies, antibody conjugates, and / or compositions thereof.

[0031] In some embodiments of the present invention, the above-mentioned nucleic acid molecules,

[0032] In the monoclonal antibody 1:

[0033] (17) Its heavy chain has the nucleotide sequence shown in SEQ ID NO:5; and

[0034] (18) Its light chain has the nucleotide sequence shown in SEQ ID NO:6; or

[0035] Nucleotide sequences obtained by modifying, substituting, deleting or adding one or more bases to the nucleotide sequences described in (19), (17) or (18);

[0036] (20) A sequence having at least 80% homology with the nucleotide sequences described in (17), (18) or (19);

[0037] (21) The complementary sequence of the nucleotide sequence described in (17), (18), (19) or (20); and / or

[0038] In the monoclonal antibody 2:

[0039] (22) Its heavy chain has the nucleotide sequence shown in SEQ ID NO:7; and

[0040] (23) Its light chain has the nucleotide sequence shown in SEQ ID NO:8; or

[0041] Nucleotide sequences obtained by modifying, substituting, deleting or adding one or more bases to the nucleotide sequences described in (24), (22) or (23);

[0042] (25) A sequence having at least 80% homology with the nucleotide sequences described in (22), (23) or (24);

[0043] (26) The complementary sequence of the nucleotide sequence described in (22), (23), (24) or (25).

[0044] The present invention also provides an expression vector comprising the above-described nucleic acid molecule.

[0045] The present invention also provides a host cell for transformation and / or transfection with the above expression vector.

[0046] The present invention also provides the use of the above-mentioned monoclonal antibody, the above-mentioned antibody-drug conjugate, the above-mentioned composition, the above-mentioned nucleic acid molecule, the above-mentioned expression vector and / or the above-mentioned host cell in any of the following:

[0047] ( ), and prepare products for detecting breast cancer; and / or

[0048] ( ), immune detection.

[0049] The present invention also provides a kit comprising: the above-described monoclonal antibody, the above-described antibody-drug conjugate, the above-described composition, the above-described nucleic acid molecule, the above-described expression vector and / or the above-described host cell, and acceptable adjuvants.

[0050] This invention provides a method for preparing a monoclonal antibody against HER-2 protein. Using HER-2 recombinant antigen as an immunogen, a hybridoma cell line is obtained through hybridoma technology. The hybridoma cell line is injected into the peritoneal cavity of mice to prepare ascites fluid, which is then purified to obtain the HER-2 monoclonal antibody. Both specific antibodies prepared by this invention have undergone rigorous clonal screening, and corresponding cell lines have been established through single-cell proliferation, exhibiting good batch-to-batch reproducibility. Experimental verification shows that the HER-2 monoclonal antibody provided by this invention demonstrates good resistance to drug interference in the detection of HER-2 protein, and can improve the accuracy of immunoassay reagents. Attached Figure Description

[0051] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below.

[0052] Figure 1 The image shows the SDS-PAGE results of the HER 20# antibody.

[0053] Figure 2 The image shows the SDS-PAGE results of the HER 16# antibody. Detailed Implementation

[0054] This invention discloses anti-HER-2 monoclonal antibodies, compositions, and their applications.

[0055] It should be understood that the expression “one or more of…” individually includes each of the objects described after the expression, as well as various different combinations of two or more of the described objects, unless otherwise understood from the context and usage. The expression “and / or” combined with three or more described objects should be understood to have the same meaning, unless otherwise understood from the context.

[0056] The terms “including,” “having,” or “containing,” including the use of their grammatical synonyms, should generally be understood as open-ended and non-restrictive, for example, not excluding other unstated elements or steps, unless otherwise specifically stated or understood from the context.

[0057] It should be understood that the order of the steps or the order in which certain actions are performed is not important as long as the invention remains operational. Furthermore, two or more steps or actions can be performed simultaneously.

[0058] The use of any and all instances or exemplary language such as “e.g.” or “including” in this document is merely intended to better illustrate the invention and is not intended to limit the scope of the invention unless the claims are made. No language in this specification should be construed as indicating that any unclaimed element is essential to the practice of the invention.

[0059] Furthermore, the numerical ranges and parameters used to define the present invention are approximate values, and the relevant values ​​in the specific embodiments have been presented as precisely as possible. However, any value inevitably contains standard deviations due to individual test methods. Therefore, unless explicitly stated otherwise, it should be understood that all ranges, quantities, values, and percentages used in this disclosure are modified with the word "approximately". Here, "approximately" generally means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a specific value or range.

[0060] This invention provides a pair of HER-2 monoclonal antibodies, wherein the amino acid sequence of the heavy chain variable region of the monoclonal antibody HER 20# used for coating is shown in SEQ ID NO:1, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO:2; the amino acid sequence of the heavy chain variable region of the monoclonal antibody HER 16# used for labeling is shown in SEQ ID NO:3, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO:4.

[0061] This invention provides a HER-2 monoclonal antibody pair, wherein the nucleotide sequence of the heavy chain variable region of the monoclonal antibody HER 20# used for coating is shown in SEQ ID NO:5, and the nucleotide sequence of the light chain variable region is shown in SEQ ID NO:6; the nucleotide sequence of the heavy chain variable region of the monoclonal antibody HER 16# used for labeling is shown in SEQ ID NO:7, and the nucleotide sequence of the light chain variable region is shown in SEQ ID NO:8.

[0062] The monoclonal antibody described in this invention specifically recognizes the HER-2 protein.

[0063] Compared with existing technologies, the monoclonal antibody provided by this invention can effectively avoid the interference of HER-2 targeted antibody drugs such as trastuzumab and pertuzumab in immune detection, improve the accuracy of detection, and thus obtain more accurate technical results.

[0064] This invention provides the application of the HER-2 monoclonal antibody pair in immunological detection products.

[0065] In some embodiments, the immunoassay product includes an immunoassay kit.

[0066] The present invention provides an immunoassay kit comprising the aforementioned HER-2 monoclonal antibody pair and acceptable excipients.

[0067] The hybridoma cells of this invention are obtained through the fusion of mouse B lymphocytes and myeloma cells, followed by multiple screenings and cloning processes. These hybridoma cells can stably secrete HER-2 monoclonal antibodies. These monoclonal antibodies exhibit high specificity, binding specifically to the HER-2 protein.

[0068] This invention provides a method for preparing the HER-2 monoclonal antibody, comprising the following steps: emulsifying the HER-2 recombinant antigen with Freund's adjuvant, immunizing mice three times, collecting spleen cells from the spleen, and hybridizing them with myeloma cells; using traditional hybridoma technology, after three rounds of screening, obtaining a series of hybridoma cells that secrete HER-2 antibodies; injecting the hybridoma cell line into the peritoneal cavity of Balb / c mice, and collecting ascites fluid 7 days later; purifying the ascites fluid using the caprylic acid-ammonium sulfate method to obtain the monoclonal antibody.

[0069] The sequence information involved in this invention is as follows:

[0070] The amino acid sequence of the variable region of the heavy chain of the monoclonal antibody HER 20# (SEQ ID NO:1):

[0071] SEQ ID NO:1:

[0072] QVQLQQSGAELMKPGASVKISCKATGYTFSNYWIEWVKQRPGHGLEWIGDILPGSDSSHNNEKFKGKVTFTADTSSNTVYMQLRSLTSEDSAVYYCTRGGGNYPYYFDFWGQGTTLTVSS;

[0073] CDR: GYTFSNYW (as shown in SEQ ID NO:9) ... ___ILPGSDSS (as shown in SEQ ID NO:10) ... ___TRGGGNYPYYFDF (as shown in SEQ ID NO:11);

[0074] The amino acid sequence of the variable region of the HER 20# light chain of the monoclonal antibody (SEQ ID NO:2):

[0075] SEQ ID NO:2:

[0076] DIVMTQSHKFMPTSVGDRVIIPCQASQDVGTAVAWYQQKSGQSPKLLIYWASTRHTGVPDRLTGSGSGTDFTLIISNVQSEDLADYFCQQYSSYPYTFGGGTKLEIKR;

[0077] CDR: QDVGTAF (as shown in SEQ ID NO:12)......___WASF (as shown in SEQ ID NO:13).......___QQYSSYPYTF (as shown in SEQ ID NO:14);

[0078] The amino acid sequence of the variable region of the monoclonal antibody HER 16# heavy chain (SEQ ID NO:3):

[0079] SEQ ID NO:3:

[0080] SDVQLQESGPGLVKPSQSLSLTCTVTGQSITSDYAWSWIRQFPGNRLEWMGYITYGGTTNYNPSLKSRISITRDTSRNQFFLQLISVTSEDTATYYCSRELRFLFVYWGQGTLVTVSA;

[0081] CDR: GQSITSDYA (as shown in SEQ ID NO:15)...___ITYGGTT (as shown in SEQ ID NO:16)...___SRELRFLFVY (as shown in SEQ ID NO:17);

[0082] The amino acid sequence of the variable region of the monoclonal antibody HER 16# light chain (SEQ ID NO:4):

[0083] SEQ ID NO:4:

[0084] DIVMSQSPSSLPVSVGEKITLSCRSSQSLLYSANQKNYLAWYQQQPGQSPKLLIYWASTRESGVPDRFTGGGSGTDFTLTISSVTAEDLAVYYCQQYYRYPYTFGGGTKLEIK;

[0085] CDR: QSLLYSANQKNY (as shown in SEQ ID NO:18) ___WAS (as shown in SEQ ID NO:19) .......___QQYYRYPYT (as shown in SEQ ID NO:20);

[0086] The nucleotide sequence of the variable region of the heavy chain of the monoclonal antibody HER 20# (SEQ ID NO:5):

[0087] SEQ ID NO:5:

[0088] CAGGTTCAGCTACAGCAGTCTGGAGCGGAGCTGATGAAGCCTGGGGCCTCAGTGAAGATTTCCTGTAAGGCTACTGGCTACACATTCAGTAACTACTGGATAGAATGGGTAAAACAGAGGCCTGGACATGGCCTTGAGTGGATTGGAGACATTTTACCTGGAAGTGATAGTTCTCACAACAATGAGAAGTTCAAGGGCAAGGTCACATTTACTGCAGATACCTCCTCCAATACAGTCTACATGCAACTCAGAAGCCTGACATCTGAAGACTCTGCCGTCTATTATTGTACAAGAGGGGGCGGTAACTACCCTTATTATTTTGACTTCTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA;

[0089] Nucleotide sequence of the light chain variable region of monoclonal antibody HER 20# (SEQ ID NO:6):

[0090] SEQ ID NO:6:

[0091] GACATTGTGATGACCCAGTCTCACAAATTCATGCCCACATCAGTAGGAGACAGGGTCATCATCCCCTGCCAGGCCAGTCAGGATGTGGGAACTGCTGTAGCCTGGTATCAACAGAAATCAGGGCAATCTCCTAAACTTCTGATTTATTGGGCATCCACCCGGCACACTGGAGTCCCTGATCGCCTCACAGGCAGTGGATCTGGGACAGATTTCACTCTCATCATTAGCAATGTGCAGTCTGAAGACTTGGCAGATTATTTCTGTCAGCAATATAGCAGTTATCCGTACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA;

[0092] Nucleotide sequence of the heavy chain variable region of monoclonal antibody HER 16# (SEQ ID NO:7):

[0093] SEQ ID NO:7:

[0094] TCTGATGTGCAACTTCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCCAGTCAATCACCAGTGATTATGCCTGGAGCTGGATCCGGCAGTTTCCAGGAAACAGACTGGAGTGGATGGGCTATATAACCTACGGTGGTACCACT AACTACAACCCATCTCTCAAAAGTCGAATCTCTATCACTCGAGACACATCCAGGAACCAGTTCTTCCTGCAGTTGATTTCTGTGACTTCTGAAGACACAGCCACATATTACTGTTCAAGAGAACTACGGTTCCTGTTTGTTTACTGGGGCCAGGGGGACTCTGGTCACTGTCTCTGCA;

[0095] The nucleotide sequence of the variable region of the monoclonal antibody HER 16# light chain (SEQ ID NO:8):

[0096] SEQ ID NO:8:

[0097] GACATTTGTGATGTCACAGTCTCCATCCTCCCTACCTGTGTCAGTTGGAGAGAAGATTACTTTGAGCTGCAGGTCCAGTCAGAGCCTTTTATAGTGCCAATCAAAAGAACTACTTGGCCTGGTACCAGCAGCAACCAGGGCAGTCTCCTAAATTGCTGATTTACTGGGC ATCCACTAGGGAGTCTGGGGTCCCTGATCGCTTCACAGGTGGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCGTGACGGCTGAAGACCTGGCAGTTTTATTACTGTCAACAATATTATAGGTATCCGTACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA.

[0098] In Examples 1 to 4 of this invention, all raw materials and reagents used can be purchased from the market.

[0099] The present invention will be further illustrated below with reference to the embodiments:

[0100] Example 1: Preparation of anti-HER-2 monoclonal antibody

[0101] 1. Mouse immunization

[0102] The HER-2 recombinant antigen was thoroughly emulsified with Freund's complete adjuvant and intraperitoneally immunized 5-week-old female Balb / c mice. The initial immunization dose was 100 μg / mouse. Subsequent immunizations were administered at 21-day intervals, with a dose of 50 μg / mouse, for a total of 3 immunizations. Approximately 10 days after the third immunization, blood was collected from the tail, and the serum titer of the mice was determined using an indirect method in 96-well plates coated with the HER-2 recombinant antigen.

[0103] 2. Hybridoma cell preparation

[0104] Select serum titers greater than 10 4 Mice underwent intrasplenic booster immunization at a dose of 100 μg per mouse. Three days after booster immunization, the mouse spleen was harvested and fused with myeloma cells NS-1. The fused cells were cultured in DMEM medium containing HAT. Approximately 6-7 days post-fusion, the cell culture supernatant was analyzed using an indirect method with 96-well plates coated with HER-2 recombinant antigen. Positive wells with an OD value not lower than 0.5 were selected for three rounds of subcloning using limiting dilution to obtain a hybridoma cell line that stably secretes anti-HER-2 antibodies.

[0105] 3. Purification of anti-HER-2 monoclonal antibody

[0106] The mouse hybridoma cells that can stably secrete anti-HER-2 antibodies were injected into the peritoneal cavity of mice, and the ascites fluid was collected and purified by octanoic acid-ammonium sulfate to obtain HER-2 monoclonal antibodies with a purity of over 85%.

[0107] Example 2: Preliminary evaluation of the anti-drug interference ability of anti-HER-2 monoclonal antibodies (antibodies screened in the above examples)

[0108] A total of 33 antibodies were screened in the HER-2 project. 96-well plates were coated with HER-2 recombinant antigen, and the competition rate between the screened monoclonal antibodies and trastuzumab and pertuzumab was measured to preliminarily evaluate the antibodies' ability to resist drug interference. The results are shown in Table 1.

[0109] Table 1 Preliminary evaluation of the anti-drug interference performance of HER-2 antibody

[0110]

[0111]

[0112] HER-2 small-sample antibodies with small cross-links with both antibody drugs were screened. HER-2 recombinant antigen was detected using a plate sandwich assay. Pairs with good reactivity and gradient were selected for drug interference evaluation. The results are shown in Table 2. The high recovery rate of the HER 20# and HRP-HER16# pairing indicates that this pairing is less affected by trastuzumab and pertuzumab.

[0113] Table 2 Crossover Rate Assessment of HER-2 Antibody-Drug Pairing

[0114]

[0115]

[0116] Example 3: Identification of HER-2 monoclonal antibody subtypes

[0117] The HER-2 recombinant antigen was diluted to 1 μg / mL and coated onto 96-well ELISA plates overnight, then blocked with 1% casein. The purified anti-HER-2 monoclonal antibody (5 mg / mL) was diluted 1:1000 with 10 mM PBS buffer. The isotype of the purified anti-HER-2 antibody was identified indirectly using a mouse monoclonal antibody isotype identification kit; the results are shown in Table 3.

[0118] Table 3. HER-2 antibody subtype identification results

[0119]

[0120] As shown in Table 3, the subtype of the anti-HER-2 monoclonal antibody is IgG1.

[0121] Example 4: Determination of the anti-drug interference performance of HER 20# and HRP-HER 16#

[0122] The kit was assembled and validated using a magnetic microparticle platform. Paramagnetic microspheres were coated with mouse anti-HER-2 monoclonal antibody HER 20# to prepare a magnetic microparticle suspension, and horseradish peroxidase (HRP)-labeled mouse anti-HER-2 monoclonal antibody HER 16# to prepare an enzyme conjugate. Six clinical samples were prepared, and 400 μg / mL of trastuzumab and pertuzumab were added to each. The samples were tested using a self-developed kit, and the results are shown in Table 4.

[0123] Table 4. Results of Anti-drug Interference Performance Tests for HER 20# and HRP-HER 16#

[0124]

[0125] The results showed that the two main pharmaceutical antibodies had no significant impact on the detection results of the magnetic microparticle platform HER 20# and HRP-HER 16#, thus improving the accuracy of the immunoassay reagent.

[0126] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A monoclonal antibody against HER-2, characterized in that, include: Monoclonal antibody 1 or monoclonal antibody 2; In the monoclonal antibody 1: (1) The amino acid sequences of the three CDR regions of its heavy chain have the amino acid sequences shown in SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:11, respectively; and (2) The amino acid sequences of the three CDR regions of its light chain have the amino acid sequences shown in SEQ ID NO:12, SEQ ID NO:13 and SEQ ID NO:14, respectively; or (3) An amino acid sequence obtained by substituting, deleting, or adding one or more amino acids to the amino acid sequence described in (1) or (2), and which has the same or similar function as the amino acid sequence shown in (1) or (2); or (4) An amino acid sequence that is at least 80% homologous to the sequence described in (1), (2) or (3); and / or In the monoclonal antibody 2: (5) The amino acid sequences of the three CDR regions of its heavy chain have the amino acid sequences shown in SEQ ID NO:15, SEQ ID NO:16 and SEQ ID NO:17, respectively; and (6) The amino acid sequences of the three CDR regions of its light chain have the amino acid sequences shown in SEQ ID NO:18, SEQ ID NO:19 and SEQ ID NO:20, respectively; or Amino acid sequences obtained by substituting, deleting, or adding one or more amino acids to the amino acid sequences described in (7), (5), or (6), and which have the same or similar function as the amino acid sequences shown in (5) or (6); or (8) An amino acid sequence that is at least 80% homologous to the sequence described in (5), (6) or (7).

2. The monoclonal antibody as described in claim 1, characterized in that, In the monoclonal antibody 1: (9) Its heavy chain variable region has the amino acid sequence shown in SEQ ID NO:1; and (10) Its light chain variable region has an amino acid sequence as shown in SEQ ID NO:2; or Amino acid sequences obtained by substituting, deleting, or adding one or more amino acids to the amino acid sequences described in (11), (9), or (10), and which have the same or similar function as the amino acid sequences shown in (9) or (10); or (12) An amino acid sequence that is at least 80% homologous to the sequence described in (9), (10) or (11); and / or In the monoclonal antibody 2: (13) Its heavy chain variable region has the amino acid sequence shown in SEQ ID NO:3; and (14) Its light chain variable region has an amino acid sequence as shown in SEQ ID NO:4; or Amino acid sequences obtained by substituting, deleting, or adding one or more amino acids to the amino acid sequences described in (15), (13), or (14), and which have the same or similar function as the amino acid sequences shown in (13) or (14); or (16) An amino acid sequence that is at least 80% homologous to the sequences described in (13), (14) or (15).

3. An antibody conjugate, characterized in that, include: The monoclonal antibody as described in claim 1 or 2, and the detectable marker.

4. A composition of monoclonal antibodies, characterized in that, include: The monoclonal antibody as described in claim 1 or 2 and / or the monoclonal antibody 1 and the monoclonal antibody 2 as described in claim 3.

5. Nucleic acid molecules encoding the monoclonal antibody as described in claim 1 or 2, the antibody conjugate as described in claim 3, and / or the composition as described in claim 4.

6. The nucleic acid molecule as described in claim 5, characterized in that, In the monoclonal antibody 1: (17) Its heavy chain has the nucleotide sequence shown in SEQ ID NO:5; and (18) Its light chain has the nucleotide sequence shown in SEQ ID NO:6; or Nucleotide sequences obtained by modifying, substituting, deleting or adding one or more bases to the nucleotide sequences described in (19), (17) or (18); (20) A sequence having at least 80% homology with the nucleotide sequences described in (17), (18) or (19); (21) The complementary sequence of the nucleotide sequence described in (17), (18), (19) or (20); and / or In the monoclonal antibody 2: (22) Its heavy chain has the nucleotide sequence shown in SEQ ID NO:7; and (23) Its light chain has the nucleotide sequence shown in SEQ ID NO:8; or Nucleotide sequences obtained by modifying, substituting, deleting or adding one or more bases to the nucleotide sequences described in (24), (22) or (23); (25) A sequence having at least 80% homology with the nucleotide sequences described in (22), (23) or (24); (26) The complementary sequence of the nucleotide sequence described in (22), (23), (24) or (25).

7. An expression vector, characterized in that, Includes the nucleic acid molecules as described in claim 6.

8. A host cell, characterized in that, Transformation and / or transfection of the expression vector as described in claim 7.

9. The use of the monoclonal antibody as described in claim 1 or 2, the antibody conjugate as described in claim 3, the composition as described in claim 4, the nucleic acid molecule as described in claim 5 or 6, the expression vector as described in claim 7, and / or the host cell as described in claim 8 in any of the following: (I) Preparing products for the detection of breast cancer; and / or (II) Immunological testing.

10. A reagent kit, characterized in that, include: The monoclonal antibody as claimed in claim 1 or 2, the antibody-drug conjugate as claimed in claim 3, the composition as claimed in claim 4, the nucleic acid molecule as claimed in claim 5 or 6, the expression vector as claimed in claim 7, and / or the host cell as claimed in claim 8, and acceptable adjuvants.

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