Method for preparing feline distemper virus monoclonal antibody by using bioreactor

Abstract

The invention discloses a method for preparing a feline distemper virus monoclonal antibody by using a bioreactor. The method comprises the following steps: step 1, carrying out subcloning on hybridoma cells by using a limited dilution method, selecting single cells into a serum-free culture medium, and carrying out suspension culture to obtain a cell strain A805 with high survival rate and strong antibody secretion ability, wherein the preservation number of the strain is CGMCC No.21902; step 2, preparing anti-feline distemper virus monoclone by using a bioreactor, wherein hybridoma cells are cultured in two stages, namely a cell growth stage and an antibody secretion stage, in the bioreactor; and step 3, harvesting the cell culture fluid as an antibody, and centrifuging to remove cell debris, thereby obtaining a finished product of the feline distemper virus monoclonal antibody. According to the method, the titer of the distemper virus monoclonal antibody is improved, the defects of a traditional ascites production antibody are overcome, and the antibody product is free of impure protein, stable in physicochemical property and small in batch-to-batch difference.

Description

technical field [0001] The invention relates to a preparation method of feline distemper virus monoclonal antibody. Background technique [0002] Feline distemper is also called feline panleukopenia, feline infectious gastroenteritis (Enteritis infectiosa felum) or feline ataxia syndrome. In 1928, Verge and Cristoforoni proved that the disease was caused by a virus. In 1930, Hammon and Ender first reported the isolation of feline distemper virus. In 1984, a strain of feline distemper virus was first isolated from natural cases in my country. [0003] Feline panleukopenia virus (Feline panleukopenia virus) is a member of the Parvoviridae family and the genus Parvovirus. In addition to infecting domestic cats, the virus can also infect animals such as tigers, leopards, minks, and raccoons. Young cats are mostly acutely ill, manifested as high fever, vomiting, diarrhea and other symptoms, and the mortality rate is as high as 90% in severe cases. Vaccination against feline di...

AI Technical Summary

Problems solved by technology

The concentration of antibodies collected by this method is high, but the operation is cumbersome, the yield is low, and large-scale production cannot be performed. Mice are susceptible to other diseases, and there will be a large amount of impurity proteins in ascites, etc.

At present, bioreactors are used to cultivate hybridoma cells, and the existing problems are mainly concentrated in the low concentration of secreted antibodies, which need to be concentrated, which greatly increases the economic cost

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