Production of Recombinant Human Monoclonal Antibody Using ras-Amplified BHK-21 Cells in a Protein-free Medium

Yuichi Inoue, Lourdes Bouzo Lopez, Seiji Kawamoto, Nobuhisa Arita, Kiichiro Teruya, Kiyohiko Seki, Masahiro Shoji, Masanori Kamei, Shuichi Hashizume, Yasumitsu Shiozawa, Hirofumi Tachibana, Hideya Ohashi, Kosei Yasumoto, Akira Nagashima, Hisashi Nakahashi, Tatsuo Suzuki, Tadaaki Imai, Kikuo Nomoto, Mitsuhiro Takenoyama, Yoshinori KatakuraSanetaka Shirahata

Research output: Contribution to journalArticlepeer-review

12 Citations (Scopus)

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 cells 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.

Original languageEnglish
Pages (from-to)811-817
Number of pages7
JournalBioscience, Biotechnology and Biochemistry
Volume60
Issue number5
DOIs
Publication statusPublished - Jan 1996

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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