PRODUCTION OF RECOMBINANT HUMAN MONOCLONAL-ANTIBODY USING RAS-AMPLIFIED BHK-21-CELLS IN A PROTEIN-FREE MEDIUM

A pas oncogene-amplified recombinant BHK-21 cell line (ras-rBHK-IgG) h as been established, and was shown to hyperproduce the recombinant IgG chimeric human monoclonal antibody (hMAb) AE6F4, which recognizes lun g cancer cells, We found that the ras-rBHK-IgG cells could be easily c ultured in a protein-free ERDF medium supplemented with iron(III) nitr ate, hydroxyethyliminodiacetic acid, and non-protein synthetic attachm ent factor as well as in a serum-free ERDF medium supplemented with in sulin, transferrin, ethanolamine, and sodium selenite, The productivit y of recombinant hMAb from the cells cultured in dishes at high cell d ensities was higher in protein-free medium than in serum-containing me dium, True high density culture of the ras-rBHK-IgG cells was done in protein-free medium using the Tecnomouse, which is a novel hollow fibe r bioreactor system, After culture for 30 days in protein-free culture , a total amount of about 14 mg of the recombinant hMAb AE6F4 was obta ined, and was shown to be reactive against lung cancer cells in tissue s.