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. 2001 Jan;35(1):9–16. doi: 10.1023/A:1008179919857

Availability of oncogene activated production system for mass production of light chain of human antibody in CHO cells

Takumi Miura 1, Yoshinori Katakura 2,✉, Perry Seto 1, Yingpei Zhang 1, Kiichiro Teruya 1, Eisaku Nishimura 3, Masatoshi Kato 3, Shuichi Hashizume 3, Sanetaka Shirahata 1
PMCID: PMC3466615  PMID: 19003276

Abstract

We previously established a ras-oncogene amplified Chinesehamster ovary (CHO) cell line, named ras clone I, as anuniversal host cell line for oncogene activated production(OAP) system to mass-produce recombinant protein by activationof the cytomegalovirus immediate early (CMV) promoter with ras protein. The λ light chain(C5λ) of human monoclonal antibody HB4C5 is expected tobe potentially useful for lung cancer targeting. We generated aC5λ hyper-producing cell line by transfecting ras cloneI with the C5λ gene expression plasmid regulated by theCMV promoter, of which productivity was 5.3 times greater thanthe hyper productive CHO cell line generated by using conventional CHO cells. Introduction of the adenovirus E1A geneinto the hyper-producing cell line derived from ras clone I resulted in further 9.5 times enhancement of the productivity,suggesting the synergistic effect of E1A and ras oncogenes on the recombinant protein production driven by the CMV promoter. In addition, intracellular accumulation of C5λ andupregulation of BiP was found in hyper-producing cell lineswhich were introduced E1A and ras oncogene. This resultsuggests that excessive intracellular accumulation ofC5λ protein, which might be caused by that the amount of produced C5λ in ER is beyond the ability of CHO cells to secrete, might signal the BiP promoter. Our data imply that ras clone I is available as a general host cell for establishing the recombinant protein hyper-producing CHOcells by the OAP system, and suggest that further mass production of recombinant proteins in the OAP system can be possible by clarifying the accurate role of upregulated BiP protein.

Keywords: cytomegalovirus promoter, HB4C5, mass production, OAP system, oncogene

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