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Cytotechnology

, Volume 33, Issue 1–3, pp 21–26 | Cite as

Properties of ras-amplified recombinant BHK-21 cells in protein-free culture

  • Yuichi InoueEmail author
  • Seiji Kawamoto
  • Masahiro Shoji
  • Shuichi Hashizume
  • Kiichiro Teruya
  • Yoshinori Katakura
  • Sanetaka Shirahata
Article

Abstract

We compared serum and protein-free cultures ofa ras-amplified recombinant BHK-21 cell line(ras-rBHK-IgG), which hyperproduces a lungcancer specific recombinant human monoclonal antibody. Ras-rBHK-IgG cells were shown to grow well, evenin protein-free medium and to be morphologicallysimilar to cells cultured in serum containing medium. However, the growth rate of ras-rBHK-IgG cellswas considerably slower in protein-free medium, whichresults in a longer maintenance period compared with cells cultured in serum containing medium. In addition, it was found that antibody production in protein-free culture had a ten times higher maximum than cells cultured in serum containing medium. On theother hand, in high density culture, using the hollowfiber bioreactor system, ras-rBHK-IgG cellscould be maintained for a month in protein-freeculture in contrast with serum culture, which onlylasted for half a month. However, the markedincrease of antibody production was not observed. A total amount of about 15 mg of the recombinantantibody, obtained in protein-free culture, was abouttwo times of that obtained in serum culture, and wasshown to be reactive to lung cancer cells in tissue. From these properties in protein-free medium, it isconcluded that protein-free culture of ras-rBHK-IgG cells is suitable for middle scaleproduction of recombinant human monoclonal antibody.

BHK-21 cells human monoclonal antibody ras oncogene synthetic attachment factor 

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Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Yuichi Inoue
    • 1
    Email author
  • Seiji Kawamoto
    • 2
  • Masahiro Shoji
    • 3
  • Shuichi Hashizume
    • 3
  • Kiichiro Teruya
    • 4
  • Yoshinori Katakura
    • 4
  • Sanetaka Shirahata
    • 4
  1. 1.Department of Biochemical Science and Technology, Faculty of AgricultureKagoshima UniversityKagoshimaJapan
  2. 2.Department of Molecular Biotechnology, Graduate School of Advanced Sciences of MatterHiroshima UniversityHigashi-HiroshimaJapan
  3. 3.Morinaga Institute of Biological ScienceYokohamaJapan
  4. 4.Graduate School of Genetic Resources TechnologyKyushu UniversityFukuokaJapan

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