论文部分内容阅读
AIM:To develop a novel process for production of HAV inVero cells grown on microcarriers in a bioreactor.METHODS:Vero cells infected with HAV strain W wereseeded at an initial density of 1×10~5 cells/mL into a 7-Lbioreactor containing Cytodex-Ⅰ microcarriers.During thestage of cell proliferation,the following conditions wereapplied:pH 7.2±0.2,temperature 37±0.2℃,dissolvedoxygen 40% of air saturation and agitation rate 40 r/min.After the stage of virus culture started,the culture conditionswere altered to pH 7.2±0.2,temperature 35±0.2℃,dissolved oxygen 25% of air saturation,agitation rate 50 r/minand perfusion of fresh medium at a flux of 20 mL/h.Duringthe course of fermentation,cell density,HAV antigen titre,glucose,lactate and ammonia levels were monitored.Acontrol experiment using conventional static culture wasconducted in the T150 flask.RESULTS:After a 28-d cultivation,cell density increasedto 14.0x10s cells/mL in the bioreactor,5.6×10~9 viable cells and4000 mL virus suspension with a titre of 1:64 were harvested.The viral antigen output per cell unit in the bioreactor was3-fold higher than that in the T150 flask.Meanwhile themetabolic mode of Veto cells did not change after the infectionwith HAV strain W.CONCLUSION:The process for production of HAV in Verocells grown on microcarriers in a bioreactor is a novel,efficient and practical way to obtain virus antigen for vaccinepurpose.This approach produces more cells and HAVantigen than the conventional static culture.With lutherimprovement,it is possible to be used for the productionof hepatitis A vaccine.
AIM: To develop a novel process for production of HAV inVero cells grown on microcarriers in a bioreactor. METHODS: Vero cells infected with HAV strain W wereseeded at an initial density of 1 × 10 ~ 5 cells / mL into a 7-Lbioreactor containing Cytodex -I microcarriers. During the cell growth, the following conditions were applied: pH 7.2 ± 0.2, temperature 37 ± 0.2 ° C, 40% of air saturation and agitation rate 40 r / min. After the stage of virus culture started, the culture Conditionswere altered to pH 7.2 ± 0.2, temperature 35 ± 0.2 ° C, dissolved oxygen 25% of air saturation, agitation rate 50 r / min and perfusion of fresh medium at a flux of 20 mL / h. Antigenic titre, glucose, lactate and ammonia levels were monitored. Control experiment using conventional static culture wasconducted in T150 flask .RESULTS: After a 28-d cultivation, cell density increasedto 14.0 × 10s cells / mL in the bioreactor, 5.6 × 10-9 viable cells and 4000 mL virus su spension with a titre of 1:64 were harvested. The viral antigen output per cell unit in the bioreactor was 3-fold higher than that in the T150 flask. Meanwhile while the themetabolic mode of Veto cells did not change after the infection with HAV strain W.CONCLUSION: The process for production of HAV in Verocells grown on microcarriers in a bioreactor is a novel, efficient and practical way to obtain virus antigen for vaccinepurpose. This approach produces more cells and HAVantigen than the conventional static culture.With lutherimprovement, it is possible to be used for the production of hepatitis A vaccine.