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교반형 막 반응기를 이용한 재조합 인간 세포의 무혈청 배지에 의한 γ-Interferon의 생산
박명식,김현규,임서규,박경유,이현용 한국산업미생물학회 1994 한국미생물·생명공학회지 Vol.22 No.4
통기축이 회전하는 배양조를 이용해 γ-IFN을 생산할 수 있는 유전자 재조합된 세포주인 HSF를 연속 배양한 결과 55(㎖/hr)의 배지 유입 속도에서 8×10exp(6)(viable cells/㎖)의 최대 세포수의 유지가 가능했으며 9000(IU/㎖)의 γ-IFN을 생산할 수 있었다. 이 같은 배양은 전반적으로 90% 이상의 생 세포수를 유지하는 우수한 공정이었다. 소비된 glucose mole 수의 80% 정도가 lactate로 일정하게 반면 glutamine의 ammonia로의 mole 전환은 배양 초기에는 극히 낮다가 배양이 안정 상태에 도달했을 때 0.4 정도의 전환율을 보였다. 특히 배양 공정이 안정상태에 도달한 경우는 lactate보다 많은 양의 ammonia가 생성돼 glutamolysis가 중요한 역할을 하고 있음을 알 수 있었다. 또한 교반에 따른 전단응력에 대해 최적 교반 속도인 50 rpm 이상의 속도에서 세포 생육은 매우 민감하게 영향을 받는 반면 최대 IFN의 비 생산 속도는 1.5×10exp(-3)(IU/cell/day)로서 γ-IFN의 생산은 다소의 감소는 있었으나 그리 크게 영향을 받지는 않았다. 전체 70일간의 배양으로 총 7.2×10exp(8)(IU)의 γ-IFN이 생산돼 약 1×10exp(7)(IU/day)의 생산성을 보여 연속 고농도 배양에 의한 상업적 생산 가능성을 확인했다. 8×10exp(6)(viable cells/㎖) of maximum cell density and 9000(IU/㎖) of γ-IFN production were obtained at 55(㎖/hr) of a perfusion rate by cultivating HSF cells using a moving membrane aeration bioreactor. This system proves to be an efficient culture process by maintaning 90% of viable cells during the whole cultivation periods. The metabolic molar quotient of glucose to lactate was 0.81 for overall ranges of glucose consumed while the evolution of ammonia was not linearly related to the consumption of glutamine. Low molar conversion ratio was observed in low consumptions of glutamine and high molar conversion ratio in high consumptions. It also shows that the glutamolysis plays important role in the steady state conditions by evolving larger quantities of ammonia than lactate. At the above of 50 rpm, which is the optimum agitation speed for this bioreactor, the cell growth was severely affected while the IFN production was less decreased, maintaing 1.5×10exp(-3)(IU/cell/day) specific IFN production rate. The cumulative γ-IFN production was 7.2×10exp(8)(IU) for 70 days of the cultivation, which yields 1×10exp(2)(IU/day) of IFN production rate. Therefore, a commercial production of γ-IFN by this culture process can be achievable by maintaining the above IFN productivity in a scaled-up culture system.
박영식,김현규,임서규,박경유,이현용,Park, Young-Shik,Kim, Hyun-Kyu,Lim, Seo-Kyu,Park, Kyung-You,Lee, Hyeon-Yong 한국미생물 · 생명공학회 1994 한국미생물·생명공학회지 Vol.22 No.4
8 X 10$^{6}$(viable cells/ml) of maximum cell density and 9000(IU/ml) of $\gamma$-IFN production were obtained at 55(ml/hr) of a perfusion rate by cultivating HSF cells using a moving membrane aeration bioreactor. This system proves to be an efficient culture process by maintaning 90% of viable cells during the whole cultivation periods. The metabolic molar quotient of glucose to lactate was 0.81 for overall ranges of glucose consumed while the evolution of ammonia was not linearly related to the consumption of glutamine. Low molar conversion ratio was observed in low consumptions of glutamine and high molar conversion ratio in high comsumptions. It also shows that the glutamolysis plays important role in the steady state conditions by evolving larger quantities of ammonia than lactate. At the above of 50 rpm, which is the optimum agitation speed for this bioreactor, the cell growth was severely affected while the IFN production was less decrea- sed, maintaing 1.5 X 10$^{-3}$(IU/cell/day) specific IFN production rate. The cumulatvie $\gamma$-IFN production was 7.2 X 10$^{8}$(IU) for 70 days of the cultivation, which yields 1 X 10$^{7}$ (IU/day) of IFN production rate. Therefore, a commercial production of $\gamma$-IFN by this culture process can be achievable by maintaining the above IFN productivity in a scaled-up culture system.