Method for improving screening efficiency of hybridoma

Abstract

The invention relates to a method aiming to improve the efficiency for screening hybridoma cells which secrete antigenic specific antibodies after cell fusion. In the method, the hybridoma cells after cell fusion are screened by using the antigen coupled solid-phase material, and the screened positive cells secreting anti-target antigen is subjected to cloning culture, so that the hybridoma cell strains of monoclonal antibody secreting the anti-target antigen can be established quickly and conveniently.

Description

[0001] Field of the invention: [0002] The invention relates to a method for screening hybridoma cells secreting specific monoclonal antibodies in the preparation of monoclonal antibodies. The establishment of the method aims to improve the screening efficiency in the hybridoma preparation process, reduce labor intensity and shorten screening time. It is one of the important foundations for the development of mass production of monoclonal antibodies. Background of the invention: [0003] The concept of monoclonal antibodies [0004] Antibodies are mainly synthesized by B lymphocytes, and each B lymphocyte has a gene for synthesizing an antibody. When the body is stimulated by an antigen, many determinants on the antigen molecule activate B cells with different genes respectively. The activated single B cell is cultured to obtain a cell group formed by division and proliferation of a single cell, which is called a monoclonal. Monoclonal cells synthesize antibodies against a...

AI Technical Summary

Problems solved by technology

Today, monoclonal antibody technology has gone through more than 30 years. During these 30 years, monoclonal antibody technology has been continuously developed and perfected day by day. The throughput and large-scale production of antibodies, this problem is how to reduce the labor cost in the process of monoclonal antibody preparation, shorten the time required for monoclonal antibody preparation, thereby reducing the cost of single monoclonal antibody production

But all these improvements are insignificant for the reduction of workload in screening

[0009] Another part of the monoclonal antibody production process that requires a lot of work is the cloning culture. The current commonly used method is the limiting dilution method. Due to the instability of the genome of the hybridoma cells, it will occur in subsequent cultures. Chromosomal loss phenomenon, even after the hybridoma cells are established, the chromosomes encoding antibodies may still be lost, making the hybridoma cells lose the ability to secrete specific antibodies. Therefore, in the initial stage of monoclonal antibody preparation, people always try to use All positive cells are cloned and cultured in order to maximize the retention of hybridoma cells secreting specific antibodies, resulting in a huge workload

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