Abstract
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.
MeSH terms
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Animals
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Antibodies, Monoclonal / biosynthesis*
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Antibodies, Monoclonal / genetics
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CHO Cells
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Cell Line, Transformed
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Cells, Cultured
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Chelating Agents / pharmacology
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Cricetinae
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Cricetulus
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Culture Media, Serum-Free
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Ethanolamine
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Ethanolamines / pharmacology
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Ferric Compounds / pharmacology
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Humans
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Imino Acids / pharmacology
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Immunoglobulin G / biosynthesis
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Immunoglobulin G / genetics
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Insulin / pharmacology
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Methacrylates / chemical synthesis
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Methacrylates / pharmacology
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Nitrates / pharmacology
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Oncogene Protein p21(ras) / genetics*
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / genetics
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / genetics
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Sodium Selenite / pharmacology
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Transferrin / pharmacology
Substances
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Antibodies, Monoclonal
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Chelating Agents
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Culture Media, Serum-Free
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Ethanolamines
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Ferric Compounds
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Imino Acids
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Immunoglobulin G
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Insulin
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Methacrylates
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Nitrates
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Recombinant Fusion Proteins
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Recombinant Proteins
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Transferrin
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Ethanolamine
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N-(2-hydroxyethyl)iminodiacetic acid
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Oncogene Protein p21(ras)
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Sodium Selenite
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ferric nitrate