Replacing factor-dependency with that for lysozyme: affordable culture of IL-6-dependent hybridoma by transfecting artificial cell surface receptor.
Cytokines and growth factors are indispensable for the propagation and maintenance of factor-dependent mammalian cells. However, cytokines are often so expensive that the use of factor-dependent cells for industrial applications such as protein production is often not practical. Based on our previous design of a binary hen egg lysozyme (HEL)-specific receptor composed of portions of the anti-HEL antibody and the erythropoietin receptor, a new pair of chimeric receptors having the intracellular domain of gp130 were made and transfected to an interleukin-6 (IL-6)-dependent hybridoma, 7TD1. The clone expressing the two new receptors showed clear HEL dose-dependent cell growth and monoclonal antibody production in both serum-based and serum-free media without IL-6. These results establish the feasibility of applying receptor design to tailor cells for the inexpensive induction of cell growth for the purpose of producing therapeutic products.
Kawahara
M
M
Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Natsume
A
A
Terada
S
S
Kato
K
K
Tsumoto
K
K
Kumagai
I
I
Miki
M
M
Mahoney
W
W
Ueda
H
H
Nagamune
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D002455:Cell Division / Q000502:physiology; D002460:Cell Line / Q000166:cytology / Q000378:metabolism; D016895:Culture Media, Serum-Free / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004526:Efficiency; D006825:Hybridomas / Q000166:cytology / Q000378:metabolism*; D015850:Interleukin-6 / Q000378:metabolism*; D051379:Mice; D009113:Muramidase / Q000378:metabolism*; D010766:Phosphorylation; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D050796:STAT3 Transcription Factor; D015534:Trans-Activators / Q000378:metabolism*; D014162:Transfection
D000911:Antibodies, Monoclonal; D016895:Culture Media, Serum-Free; D004268:DNA-Binding Proteins; D015850:Interleukin-6; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D050796:STAT3 Transcription Factor; C494087:Stat3 protein, mouse; D015534:Trans-Activators; C421864:hen egg lysozyme; D009113:Muramidase