500L生物反应器生产人二倍体细胞狂犬病疫苗工艺的研究

Manufacture of Human Diploid Cell Rabies Vaccine Using 500 L Bioreactors

目的通过500 L生物反应器培养人二倍体细胞,接种感染PM株狂犬病毒,监测人二倍体细胞生长情况和病毒表达情况,确定一套可行的疫苗生产放大工艺。方法采用一定细胞传代接种比例培养人二倍体细胞、同时按照一定的病毒接种比例感染细胞,监测细胞病毒培养过程中的细胞密度及病毒表达情况,从而确定生产工艺的可行性。结果经过50 L和500 L生物反应器培养MRC-5细胞,接种感染狂犬病毒PM株毒种后,细胞生长良好,细胞密度、病毒滴度表达水平及表达持续时间均能达到疫苗生产要求。结论用500 L生物反应器培养MRC-5细胞,生产人二倍体细胞狂犬病疫苗,该生产放大工艺是可行的。

Objective To infect human diploid cells incubated in 500 L bioreactors with PM strain of rabies virus and monitor diploid cell growth and virus expression so as to develop a feasible vaccine production scale- up process. Methods Human diploid cells were incubated in 500 L bioreactors at a specified passage ratio and then infected at a specified virus inoculation ratio; diploid density and virus expression during incubation were monitored so as to determine feasibility of the manufacturing process. Results Following inoculation of PM strain of rabies virus to MRC- 5 cells incubated in 50 L and 500 L bioreactors,the cells grew well; cell density,virus titer expression level and duration could all meet vaccine manufacture requirements. Conclusion The production scale- up process of incubating MRC- 5 cells using 500 L bioreactors to manufacture diploid cell rabies vaccine is feasible.