Semi-solid medium and its preparation method and hybridoma screening method

Abstract

The invention relates to a semisolid culturing medium. The semisolid culturing medium comprises a methyl cellulose solution, a 2*1640 culturing medium, an L-glutamine solution, a beta-mercaptoethanol solution, fetal bovine serum, a penicillin and streptomycin mixed solution, an HAT solution and an antigen-coated nano-gold solution. The semisolid culturing medium can be directly used for culturing and screening hybridoma cells, can reduce the total subcloning frequency of a culturing process to greatly reduce the labor intensity, and also can make the growth spaces of single cell lines independent to reduce the contact inhibition of clones; and the introduction of antigen-coated nano-gold particles realizes one-step of screening of monoclonal cell lines secreting target antibodies in order to save the monoclonal screening time. The invention also relates to a preparation method of the semisolid culturing medium, and a hybridoma cell screening method using the semisolid culturing medium.

Description

Technical field [0001] The invention relates to the field of cell engineering, in particular to a semi-solid culture medium, a preparation method thereof, and a hybridoma cell screening method. Background technique [0002] Antibodies are mainly synthesized by B lymphocytes, and each B lymphocyte contains genetic genes that synthesize an antibody. When the body is stimulated, many antigenic determinants on antigen molecules activate B lymphocytes with different genes. The activated single B lymphocytes are cultured to obtain a cell population formed by the division and proliferation of single cells, which is called monoclonal. Monoclonal cells synthesize antibodies against an antigenic determinant, namely monoclonal antibodies (abbreviated as monoclonal antibodies). [0003] In 1975, the German scholar Kohler and the British scholar Milstein fused mouse spleen cells immunized with sheep red blood cells from the nucleus of mouse myeloma cells in vitro to form hybridoma cells, whic...

AI Technical Summary

Problems solved by technology

Today, monoclonal antibody technology is being continuously improved, but there is a problem that has been puzzling the researchers of monoclonal antibody preparation and restricting the throughput and large-scale production of monoclonal antibody, that 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 a single monoclonal antibody preparation

Therefore, in the initial stage of monoclonal antibody preparation, people always clone and culture as many positive cells as possible in order to retain the hybridoma cells secreting specific antibodies to the greatest extent, resulting in a huge workload

The limited dilution method is not only labor-intensive and takes a long time, but also requires a lot of experience in cloning screening, strictly controlling the time of subcloning, and the operation is cumbersome.

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