CN-121824753-B - Anti-GATA 3 recombinant rabbit monoclonal antibody and application thereof

Abstract

The invention belongs to the technical field of immunochemistry, and particularly relates to an anti-GATA 3 recombinant rabbit monoclonal antibody and application thereof, wherein the anti-GATA 3 recombinant rabbit monoclonal antibody comprises a heavy chain variable region and a light chain variable region, the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 8, and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 9. The invention also relates to a nucleotide sequence SEQ ID NO. 6 and SEQ ID NO. 7 of the anti-GATA 3 recombinant rabbit monoclonal antibody, a nucleic acid molecule, an expression vector or recombinant plasmid, a host cell, a preparation method of the antibody, application of the antibody in a GATA3 protein detection method or device and the like. The GATA3 recombinant rabbit monoclonal antibody has the characteristics of good specificity, strong positive signals and the like, is easier to score in IHC staining, and is more accurate for detecting and distinguishing cancers.

Inventors

• Xiao Muzhang
• OuYang Lingzi
• WU SHANGJUN
• ZHANG DONGXU

Assignees

• 苏州百道医疗科技有限公司
• 中南大学湘雅医院
• 湘雅常德医院

Dates

Publication Date

20260508

Application Date

20260311

Claims (10)

1. 1. The anti-GATA 3 recombinant rabbit monoclonal antibody is characterized by comprising a heavy chain variable region and a light chain variable region, wherein the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 8, and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 9.
2. 2. A coding gene for the anti-GATA 3 recombinant rabbit monoclonal antibody of claim 1.
3. 3. The coding gene according to claim 2, comprising a DNA sequence as shown in SEQ ID NO. 6 for encoding the heavy chain variable region of the anti-GATA 3 recombinant rabbit monoclonal antibody and a DNA sequence as shown in SEQ ID NO. 7 for encoding the light chain variable region of the anti-GATA 3 recombinant rabbit monoclonal antibody.
4. 4. A nucleic acid molecule comprising the coding gene of claim 2 or 3.
5. 5. An expression vector comprising the nucleic acid molecule of claim 4.
6. 6. A host cell transformed or transfected with the expression vector of claim 5.
7. 7. A preparation method of an anti-GATA 3 recombinant rabbit monoclonal antibody is characterized in that the expression vector of claim 5 is adopted to transform or transfect host cells, the transformed or transfected cells are cultured, cell supernatant is collected and purified, and the anti-GATA 3 recombinant rabbit monoclonal antibody is obtained.
8. 8. Use of an anti-GATA 3 recombinant rabbit monoclonal antibody according to claim 1, a coding gene according to claim 2 or 3, a nucleic acid molecule according to claim 4, an expression vector according to claim 5, a host cell according to claim 6 for the preparation of a GATA3 assay device.
9. 9. A GATA3 assay kit comprising the anti-GATA 3 recombinant rabbit monoclonal antibody of claim 1 and an immunohistochemical detection reagent.
10. 10. The GATA3 detection kit of claim 9 comprising the anti-GATA 3 recombinant rabbit monoclonal antibody, horseradish peroxidase-labeled secondary antibody, ethylenediamine tetraacetic acid repair solution, catalase blocking solution, 3 '-diaminobenzidine concentrate, 3' -diaminobenzidine buffer, hematoxylin and bluing solution of claim 1.

Description

Anti-GATA 3 recombinant rabbit monoclonal antibody and application thereof Technical Field The invention belongs to the technical field of immunochemistry, and particularly relates to an anti-GATA 3 recombinant rabbit monoclonal antibody and application thereof, in particular to application in immunohistochemical detection. Background GATA3 (GATA binding protein 3) is a key member of the GATA transcription factor family, a highly conserved zinc finger structural transcription factor with a molecular weight of about 48kDa and is primarily located in the nucleus. The core function of the GATA3 protein is to regulate gene transcription by combining with a target gene regulation region, so as to participate in cell development, differentiation fate determination and tissue phenotype maintenance, and the function of the GATA3 protein has obvious tissue specificity. GATA3 expression has extremely strict tissue specificity, and its core is highly expressed in two major systems, namely, epithelial tissue represented by mammary gland, kidney, etc., and hematopoietic immune cells represented by T lymphocytes, NK cells, etc. Whereas in non-target tissues GATA3 exhibits a low or no expression state. The strict space-time expression specificity endows GATA3 with extremely high pathological diagnosis value, so that the GATA3 becomes a core immunohistochemical marker for clinically distinguishing and diagnosing epithelial tumors (especially breast cancer and the like), and is also used as an auxiliary index for prognosis evaluation of partial tumors. The abnormal expression (high expression/deletion) of GATA3 protein is closely related to tumorigenesis and development, and the nuclear expression characteristics and tissue specificity of the GATA3 protein make the GATA3 protein become a core marker for differential diagnosis of epithelia tumors in clinical pathological diagnosis, and are also auxiliary indexes for prognosis evaluation of partial tumors. At present, detection of GATA3 proteins mainly depends on immunohistochemistry, but the existing anti-GATA 3 antibodies have limited capture capability on low-abundance targets, and weak positive samples are frequently missed due to insufficient sensitivity, so that false negative results are generated. This defect affects in particular the accurate identification of early tumors, poorly differentiated tumors and partial metastases, limiting the clinical efficacy of GATA3 as a diagnostic marker. Therefore, the anti-GATA 3 antibody reagent which can efficiently identify the GATA3 protein in the tissue sample, has higher sensitivity and stronger specificity and can adapt to the processing conditions of clinical conventional samples is developed, and has important practical significance and clinical application value for improving the diagnosis accuracy of epithelial tumors and the like and promoting the deep development of the related foundation and clinical research of GATA 3. Disclosure of Invention First, the technical problem to be solved In view of the above-mentioned shortcomings and disadvantages of the prior art, the present invention provides an anti-GATA 3 recombinant rabbit monoclonal antibody which has a wide application and can accurately recognize GATA3 expression, and its application. The invention also relates to a nucleotide sequence, a recombinant plasmid or an expression vector for encoding the anti-GATA 3 recombinant rabbit monoclonal antibody, a preparation method, application of the anti-GATA 3 recombinant rabbit monoclonal antibody in a GATA3 protein detection method or device and the like. (II) technical scheme In order to achieve the above purpose, the main technical scheme adopted by the invention comprises the following steps: In a first aspect, the invention provides an anti-GATA 3 recombinant rabbit monoclonal antibody, which comprises a heavy chain variable region and a light chain variable region, wherein the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 8, and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 9. The anti-GATA 3 recombinant rabbit monoclonal antibody (GATA 3 rabbit-derived antibody) can be used for immunohistochemical detection, and can identify and detect the expression of GATA3 protein on tumor cells or immune cells with high specificity and high sensitivity. The anti-GATA 3 monoclonal antibody is obtained by recombinant expression of mammalian cells. Specifically, the anti-GATA 3 recombinant rabbit monoclonal antibody provided by the invention is produced by eukaryotic expression of 293 cells through rabbit hybridoma fusion screening. In the preparation of the anti-GATA 3 monoclonal antibody, an antigen for immunizing rabbits (New Zealand white rabbits) is a synthetic polypeptide, the amino acid sequence of which is shown as SEQ ID NO. 1, and the synthetic polypeptide is obtained by artificial chemical synthesis. After the rabbit is immunized, a positive hybridom