应用生物反应器悬浮培养水貂犬瘟热疫苗株工艺的优化  

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

作　　者：盛程程 黄东东 李文娟 陈立志 SHENG Cheng-cheng;HUANG Dong-dong;LI Wen-juan;CHEN Li-zhi(College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,Jilin Province,China;不详)

机构地区：[1]吉林农业大学动物科学技术学院,吉林长春130118 [2]吉林特研生物技术有限责任公司,吉林长春130112

出　　处：《中国生物制品学杂志》2020年第2期194-196,202,共4页Chinese Journal of Biologicals

基　　金：吉林省科技攻关计划。

摘　　要：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。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.

关 键 词：生物反应器 水貂犬瘟热疫苗 CDV3-CL株 微载体 悬浮培养 

分 类 号：R392-33[医药卫生—免疫学]