应用生物反应器悬浮培养水貂犬瘟热疫苗株工艺的优化

目的对应用生物反应器悬浮培养水貂犬瘟热疫苗株的工艺进行优化。方法采用控制变量法对水貂犬瘟热疫苗株(CVD3-CL株)在生物反应器中微载体悬浮培养的相关参数[培养温度(30、33、37℃)、病毒感染复数(MOI,0. 01、0. 03、0. 05、0. 07)及收毒时间]进行优化,并对制备的疫苗进行动物安全性和攻毒保护试验。结果在5 g微载体、1 L培养体系中,温度设定为33℃,MOI设为0. 03时,更有利于病毒的复制增殖,病毒滴度可在接毒后108 h达峰值,为10^6.7 TCID50;制备的疫苗安全性良好,可有效免疫水貂,保护率为100%。结论优化了应用生物反应器悬浮培养水貂犬瘟热弱毒疫苗株的工艺,为水貂犬瘟热弱毒疫苗的大规模培养提供了参考。

Optimization of procedure for suspension culture of mink canine distemper vaccine strain by using bioreactor

Objective To optimize the procedure for suspension culture of mink distemper vaccine strain by using bioreactor. Methods The parameters for suspension culture of mink distemper vaccine strain CVD3-CL,including temperature(30,33 and 37 ℃),MOI(0. 01,0. 03,0. 05 and 0. 07) and harvest time,were optimized by control variate method,and the prepared vaccine was subjected to safety test and challenge protection test in animals. Results Suspension culture on 5 g microcarriers in 1 L cultivation system at a temperature of 33 ℃ and a MOI of 0. 07 was more beneficial to the virus replication and proliferation. The virus titer reached a peak value of 10^6.7 TCID50108 h after inoculation.The prepared vaccine showed high safety,of which the protective rate to mink was 100%. Conclusion The procedure for suspension culture of mink distemper vaccine strain by using bioreactor was optimized,which provided a reference for the large-scale culture of virus strain for production of live attenuated mink canine distemper vaccine.