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KMID : 0545120040140040810
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Journal of Microbiology and Biotechnology
2004 Volume.14 No. 4 p.810 ~ p.815
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High-Level Expression of Recombinant Human Interleukin-2 in Chinese Hamster Ovary Cells Using the Expression System Containing Transcription Terminator
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Kim EJ
Kim DJ/Hwang HY/Yoon JS/Yoon Y/Baek KH
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Abstract
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Many biological properties and the clinical potential of human interleukin-2 (hIL-2) draw much attention to its high-level expression in mammalian cells. Recombinant human IL-2 (rhIL-2) was expressed in Chinese hamster ovary (CHO) cells using the recently developed expression system which confers position-independent expression. Stable CHO cell lines carrying several hundred amplified copies of the rhIL-2 gene were easily obtained and rhIL-2 was expressed at high levels after selection with increasing concentrations of methotrexate. Interestingly the insertion of the transcription terminator of the human gastrin gene into the downstream region of the gene for rhIL-2 considerably increased rhIL-2 expression. Using the expression system with the transcription terminator it was possible to get a CHO cell line expressing the rhIL-2 at a very high level about 11.4 μg/106 cell/day which is about 6 times higher than that previously reported. The biological activity of the rhIL-2 protein purified from the cell line was also confirmed by the cell proliferation assay.
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