High-level production of thrombopoietin by recombinant chinese hamster ovary cells = 재조합 CHO 세포에 의한 혈소판증식인자(Thrombopoietin)의 고농도 생산

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When two recombinant Chinese hamster ovary (rCHO) cell lines producing human thrombopoietin (hTPO), CHO/dhfr-B22-4 and W44, were cultivated in hyperosmolar media resulting from NaCl addition, their specific foreign protein productivity increased with medium osmolality. However, due to a simultaneous suppression of cell growth at elevated osmolality, no enhancement in the maximum foreign protein titer was made in batch cultures. To test the feasibility of using glycine betaine, known as a strong osmoprotective compound, for improved foreign protein production in hyperosmotic rCHO cell cultures, hyperosmotic batch cultures were carried out in the presence of 15 mM glycine betaine. Glycine betaine was found to have a strong osmoprotective effect on both rCHO cell lines. Inclusion of 15 mM glycine betaine in hyperosmolar medium enabled rCHO cell lines to grow at 571-573 mOsm/kg where they could not grow in the absence of glycine betaine. However, effect of glycine betaine inclusion in hyperosmolar medium on foreign protein production differed between rCHO cell lines. CHO/dhfr-B22-4 cells retained enhanced specific hTPO productivity ($q_{hTPO}$) in the presence of glycine betaine, and thereby, the maximum hTPO titer obtained at 573 mOsm/kg was increased by 72% over that obtained in the control culture with physiological osmolality (292 mOsm/kg). On the other hand, enhanced $q_{hTPO}$ of W44 at elevated osmolality decreased significantly in the presence of glycine betaine at a cost of the recovery of cell growth. As a result, the maximum antibody titer at 571 mOsm/kg was similar to that obtained in the control culture with physiological osmolality. Taken together, efficacy of the simultaneous use of hyperosmotic pressure and glycine betaine as a means to improve foreign protein production was variable among different rCHO clones. When parental rCHO clones producing hTPO were subjected to higher methotrexate (MTX) level, clonal variations in regard to cell growth we...
Kim, Sun-Changresearcher김선창researcher
한국과학기술원 : 생물과학과,
Issue Date
158694/325007 / 000983154

학위논문(석사) - 한국과학기술원 : 생물과학과, 2000.2, [ ix, 83 p. ]

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