复制型重组痘苗活载体疫苗生产工艺研究

Optimization of Production Procedure of Replicating Recombinant Live Vaccinia Virus Vector-based Vaccine

目的研究复制型重组痘苗活载体疫苗的生产工艺。方法通过在病毒-细胞比(MOI)、不同培养时间、培养温度、碳酸氢钠含量的条件下培养复制型重组痘苗病毒,确定适用于重组痘苗活载体疫苗的生产工艺。以确定的生产工艺制备3批疫苗,并进行各项检定。结果以MOI为1∶750,培养液中加入7.5%碳酸氢钠的比例为1.0%,培养温度为37℃,培养时间为72 h左右,获得疫苗的病毒滴度最高。按此生产工艺生产的3批疫苗,病毒滴度均在107PFU/ml以上,经检定各项指标均合格。结论确定了复制型重组痘苗活载体疫苗的生产工艺。

Objective To optimize the production procedure of replicating recombinant live vaccinia virus vector-based vaccine. Methods Culture replicating recombinant live vaccinia virus vector, in which HIV antigen gene was inserted,at various MOIs, temperatures and sodium bicarbonate concentrations for various hours to optimize the production prrocedure, Throe batches of vaccine were prepared by the optimized procedure for control tests. Results The production procedure was opthnized as follows ;inoculate recombinant virus into ceils at a MOI of 1：750,add 7.5% sodium bicarbonate into medium to a final concentration of 1.0% and incubate at 37℃ for about 72 h. All the 3 batches of vaccines prepared by the procedure reached virus titers of more than 107 PFU/ml and were qualified in control tests. Conclusion The production procedure of replication recombinant live vaccinia virus vector-based vaccine was optimized.