Method for treating contaminated hybridoma cells through mouse model

Abstract

The invention discloses a method for treating contaminated hybridoma cells through a mouse model. The method comprises the steps that abdominal pretreatment is conducted on a 10-week-old mouse to build the mouse model; a contaminated hybridoma cell strain secreting monoclonal antibodies are suspended and centrifuged, a culture medium is removed, cell counting is conducted under an inverted microscope, and after the cell density is regulated to be 8*10<5> / cm<2>, the cells are injected into the abdominal cavity of the built mouse model; ascites is collected 5 days later, the ascites collected for the first time is centrifuged, bottom precipitates are retained, resuspended with normal saline, washed and then centrifuged, and bottom white precipitates are retained; the white precipitates are resuspended and cultured with a prepared DMEM culture medium, the growth state of the hybridoma cells is observed, and when the hybridoma cells grow to a logarithmic growth phase, supernatant liquid is taken, and the titer of the supernatant liquid is detected; the hybridoma cells meeting the sterilization requirement are pretreated and then cryopreserved for breed conversation. The method has the advantages of being simple, convenient to operate and high in safety.

Description

technical field [0001] The invention relates to the preparation of monoclonal antibodies, in particular to a method for treating polluting hybridoma cells using a mouse model. Background technique [0002] In 1975, Kohler and Milstein established the hybridoma technology, which fused mouse myeloma cells with mouse splenocytes producing sheep red blood cells. The hybridoma cells formed could not only produce antibodies but also divide and reproduce, and the antibodies produced could only recognize One type of antigenic determinant, known as a monoclonal antibody. Monoclonal antibody is a major research hotspot in future therapeutics. It is reported that monoclonal antibody drugs will become the main theme of biomedical research. So far, thousands of monoclonal antibodies have been developed in the world, and are widely used in the fields of biology and medicine, and play an important role in the diagnosis and treatment of disease pathogenesis. With the continuous deepening ...

AI Technical Summary

Problems solved by technology

At present, for cell lines with a large number of seed banks, strong alkali can be used for direct disinfection treatment (generally 0.2molNaOH); for cell lines where pollution sources can be clearly detected, antibiotics (generally penicillin, etc.) can be added to inhibit the growth of pollution sources, and Cleaning is carried out by frequent medium changes, but the success rate is not high; and how to sterilize some rare hybridoma cell lines that are very important to the entire experiment after contamination, there is no relatively mature method for the time being