High-level production of human thrombopoietin by anti-apoptotic CHO cells in serum-free suspension culture = 무혈청 부유배양에서 항 세포예정사 CHO 세포에 의한 인간 혈소판증식인자의 고농도 생산

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For the high-level production of human thrombopoietin (hTPO) in serum-free suspension culture of recombinant Chinese hamster ovary (rCHO) cells, the medium was optimized by the use of medium additives and CHO cells were engineered by the regulation of gene expression. The performance of a serum-free medium was enhanced by yeast hydrolysate addition. Sodium butyrate (NaBu) addition significantly increased both the specific and volumetric hTPO production, although it decreased the cell viability by apoptosis in a dose-dependent manner. However, NaBu deteriorated the quality of hTPO, resulting from increased heterogeneity, reduced acidic hTPO isoforms, reduced a(2→3) sialylation, and decreased in vivo biological activity. To overcome detrimental effects of NaBu on apoptotic cell death and hTPO quality, I overexpressed Bcl-2 protein or downregulated caspases in rCHO cells producing hTPO. Bcl-2 overexpression in rCHO cells and NaBu addition in serum-free suspension culture can be an effective means to enhance the production of highly glycosylated protein such as hTPO. Compared to serum-free suspension culture of control cells without Bcl-2 overexpression (R-neo cells) and NaBu addition, a more than 10-fold increase in the maximum hTPO concentration was obtained in serum-free suspension culture of cells with Bcl-2 overexpression (R-bcl2-14 cells) and 3 mM NaBu addition.
Advisors
Lee, Gyun-Minresearcher이균민researcher
Description
한국과학기술원 : 생명과학과,
Publisher
한국과학기술원
Issue Date
2005
Identifier
245032/325007  / 020015151
Language
eng
Description

학위논문(박사) - 한국과학기술원 : 생명과학과, 2005.2, [ viii, 154 p. ]

Keywords

Bcl-2; thrombopoietin; CHO cells; apoptosis; caspase; 카스파제; Bcl-2; 혈소판증식인자; CHO 세포; 세포예정사

URI
http://hdl.handle.net/10203/27594
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=245032&flag=dissertation
Appears in Collection
BS-Theses_Ph.D.(박사논문)
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