Latex semisolid culturing medium, preparation method thereof, and hybridoma cell screening method

A semi-solid medium and culture medium technology, applied in the field of cell engineering, can solve the problems of long cycle, huge workload, restriction of monoclonal antibody flux and large-scale production, etc.

Active Publication Date: 2014-07-02
SHENZHEN INST OF ADVANCED TECH
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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 preparat...

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  • Latex semisolid culturing medium, preparation method thereof, and hybridoma cell screening method

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preparation example Construction

[0044] Such as figure 1 Shown, the preparation method of the semi-solid culture medium of one embodiment, comprises the steps:

[0045] Step S110, preparing respectively a methylcellulose solution with a concentration of 0.4 g / ml, a 2×1640 medium with a pH of 7.0 to 7.4, an L-glutamine solution with a concentration of 1 mol / l, and a β-glutamine solution with a concentration of 1 mmol / l. - mercaptoethanol solution.

[0046] Step S120, diluting the antigen to 100 μg / ml with 0.1 mmol / l phosphate buffer, taking 10% polystyrene latex and diluting it with 0.1 mmol / l phosphate buffer at a ratio of 1:5, Then the diluted polystyrene latex solution was mixed with the diluted antigen solution in equal volumes, and allowed to stand at 37° C. for 1 hour to obtain a latex antigen particle solution.

[0047] Step S130, preparing latex semi-solid medium: adding the 2×1640 medium, the L-glutamine solution, the β-mercaptoethanol solution, fetal calf serum, 10,000 units / ml of penicillin and 1...

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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 latex antigen particles. 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. 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 latex semi-solid 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 the genetic gene for synthesizing an antibody. When the body is stimulated, many antigenic determinants on the antigen molecule respectively activate B lymphocytes with different genes. The activated single B lymphocytes are cultured to obtain a cell group formed by the division and proliferation of single cells, which is called monoclonal. Monoclonal cells synthesize antibodies against an antigenic determinant, namely monoclonal antibodies (referred to as monoclonal antibodies). [0003] In 1975, German scholar Kohler and British scholar Milstein fused the mouse splenocytes with the nucleus of mouse myeloma cells immunized with sheep red blood cells in vitro to form hybrido...

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Application Information

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IPC IPC(8): C12N5/20G01N33/569
Inventor 万晓春刘婕金言
Owner SHENZHEN INST OF ADVANCED TECH
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