On December 25, 2011, Molecular Biotechnology online published the Zhou Paul research group of the Shanghai Pasteur Institute of Chinese Academy of Sciences titled "Perfusion culture with wave bioreactor to stably transfer human monoclonal antibody gene Drosophila cell line high-yield human list "Cloning antibodies" article. This is the first report on the production of human monoclonal antibodies by perfusion cultivation of Drosophila S2 cells stably transformed with human monoclonal antibody genes using a wave bioreactor.
Therapeutic human monoclonal antibodies have grown into a multi-billion dollar industry, so it is very important to establish a stable culture condition to meet the large-scale expression of human monoclonal antibodies. The wave bioreactor was used for protein expression in mammals and insect cells in the late 1990s, but the wave bioreactor was used to perfuse the cultured Drosophila stably transformed cells to express human This is the first time cloned antibody protein.
In this study, under the guidance of Professor Zhou Paul, Wang Lulan, a research assistant at the Shanghai Pasteur Institute, and Hu Hongxing, a doctoral student, cloned the human monoclonal antibody gene against hemagglutinin of highly pathogenic avian influenza virus and constructed Into the Drosophila inducible expression vector, the Drosophila S2 cell line was stably transformed, and a single cell clone that highly expressed human monoclonal antibodies was obtained by limiting dilution. Monoclonal cell lines stably transformed with human monoclonal antibodies were used for comparative cultivation in perfused and non-perfused bioreactors, and it was found that the perfusion culture method was significantly better than the latter in obtaining the number of cells and the yield of expressed antibody protein. And the antibodies produced by these two methods have neutralizing activity.
These results indicate that the wave bioreactor is a very good method for perfusion culture of S2 stable transfected cell lines for large-scale protein expression.
The research was completed in collaboration with Dr. Yang Jianjun of the Fast Trak Center of GE ’s R & D Department, and was supported by the National Natural Science Foundation of China and the Li Ka Shing Foundation.
Comparison of cell growth rate and antibody production in perfusion and non-perfusion culture
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