Production of recombinant human monoclonal antibody using ras-amplified BHK-21 cells in a protein-free medium.
Y Inoue, L B López, S Kawamoto, N Arita, K Teruya, K Seki, M Shoji, M Kamei, S Hashizume, Y Shiozawa, H Tachibana, H Ohashi, K Yasumoto, A Nagashima, H Nakahashi, T Suzuki, T Imai, K Nomoto, M Takenoyama, Y Katakura, S Shirahata
文献索引:Biosci. Biotechnol. Biochem. 60(5) , 811-7, (1996)
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摘要
A ras oncogene-amplified recombinant BHK-21 cell line (ras-rBHK-IgG) has been established, and was shown to hyperproduce the recombinant IgG chimeric human monoclonal antibody (hMAb) AE6F4, which recognizes lung cancer cells. We found that the ras-rBHK-IgG cell could be easily cultured in a protein-free ERDF medium supplemented with iron(III) nitrate, hydroxyethyliminodiacetic acid, and non-protein synthetic attachment factor as well as in a serum-free ERDF medium supplemented with insulin, transferrin, ethanolamine, and sodium selenite. The productivity of recombinant hMAb from the cells cultured in dishes at high cell densities was higher in protein-free medium than in serum-containing medium. True high density culture of the ras-rBHK-IgG cells was done in protein-free medium using the Tecnomouse, which is a novel hollow fiber bioreactor system. After culture for 30 days in protein-free culture, a total amount of about 14 mg of the recombinant hMAb AE6F4 was obtained, and was shown to be reactive against lung cancer cells in tissues.
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