Preparation method of beta2-microglobulin monoclonal antibody

A monoclonal antibody and microglobulin technology, applied in immunoglobulin, anti-animal/human immunoglobulin, chemical instruments and methods, etc., can solve the problem that it is difficult to obtain positive monoclonal antibodies and mice are difficult to obtain positive Monoclonal antibody and other issues, to achieve the effect of increasing the positive rate of monoclonal antibody
CN110642949AActive Publication Date: 2020-01-03上海钹乐诗生物技术有限公司
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
上海钹乐诗生物技术有限公司
Publication Date
2020-01-03

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Abstract

The invention discloses a preparation method of a beta2-microglobulin monoclonal antibody. The method comprises the following steps: firstly, screening out two polypeptide epitope sequences with relatively strong immunogenicity as a first antigen epitope and a second antigen epitope respectively; then respectively connecting one cysteine to each carbon-terminal amino acid of the first antigen epitope and the second antigen epitope, and coupling sulfydryl of the cysteine with primary amino of keyhole limpet hemocyanin through an SMCC coupling method so as to obtain an in-vitro synthesized beta2-MG double-epitope polypeptide antigen; then carrying out conventional immunization on the mouse by utilizing the double-epitope polypeptide antigen, and separating out splenic lymphocytes and myelomacells to carry out cell fusion; and finally, screening out positive monoclonal cells, and extracting monoclonal antibody through intraperitoneal injection and ascites recovery. By improving traditional single-epitope antigen immunization methods, the method optimizes the immunogenicity of the antigen, shortens the immunization period, and improves the serum titer in the immunization stage, thereby improving the obtaining efficiency of the positive monoclonal antibody against beta2-MG.
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Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a method for preparing a β2-microglobulin monoclonal antibody. Background technique

[0002] β2 microglobulin, referred to as β2-MG, is a small molecular globulin secreted from lymphocytes, platelets and polymorphonuclear leukocytes. It is a single-chain linear polypeptide composed of 99 amino acids with a molecular weight of 11,800. β2-MG is a component of HLA-class I molecules. The concentration of β2-MG in normal human serum is in the range of 0.5-2.0mg / L. The only organ that excretes β2-MG out of the body is the kidney. When renal insufficiency occurs When β2-MG is excreted, the concentration of β2-MG in the patient's serum is 60 times that of normal people. When β2-MG is excessively deposited in the tissue, it is easy to induce β2-MG-related amyloidosis. Studies have found that the level of β2-MG in the serum is significantly negatively correlated with the glomerular filtration...

Claims

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