生物反应器悬浮培养禽偏肺病毒制备弱毒疫苗工艺的优化  

Optimization of procedure for preparation of attenuated vaccine by suspension culture of avian metapneumovirus in bioreactor

作　　者：蒋天华 周庆丰 严专强 周展[1] JIANG Tian-hua;ZHOU Qing-feng;YAN Zhuan-qiang;ZHOU Zhan(College of Pharmaceutical Sciences,Zhejiang University Hangzhou 310058,Zhejiang Province,China;不详)

机构地区：[1]浙江大学药学院,浙江杭州310058 [2]温氏食品集团股份有限公司,广东云浮527439

出　　处：《中国生物制品学杂志》2022年第3期316-320,共5页Chinese Journal of Biologicals

基　　金：广东省基础与应用基础研究基金(2019B1515210008)。

摘　　要：目的对生物反应器悬浮培养禽偏肺病毒(avian metapneumovirus,aMPV)制备弱毒疫苗的工艺进行优化。方法采用分批培养的方法优化悬浮BHK-21细胞在生物反应器内培养及aMPV增殖工艺,并对制备的疫苗进行动物安全性及攻毒保护试验。结果采用1.0×10^(6)cells/mL的初始接种密度培养BHK-21悬浮细胞,细胞能快速度过延滞期进入对数生长期,培养72 h细胞密度可达1.6×10^(7)cells/mL。调整细胞密度为3.0×10^(6)cells/mL,按照感染复数MOI=0.1的剂量接种病毒,病毒含量可达10^(8.67)TCID_(50)/mL。制备的疫苗安全性良好,可有效保护番鸭,保护率为100%。结论优化了生物反应器培养aMPV的工艺,可获得较高的病毒效价,为aMPV疫苗的规模化生产提供了参考。Objective To optimize the procedure for preparation of attenuated vaccien by suspension culture of avian metapneumovirus(aMPV)in bioreactor.Methods The procedure for suspension culture of BHK-21 cells in bioreactor and propagation of aMPV was optimized by batch-wise culture,and the prepared vaccine was subjected to safety test and challenge protection test in animals.Results The BHK-21 cells at an initial concentration of 1.0×10^(6) cells/mL in suspension culture entered the logarithmic growth phase immediately,of which the density reached 1.6×10^(7) cells/mL72 h later.After the cells were adjusted to a density of 3.0×10^(6) cells/mL and inoculated with aMPV at a MOI of 0.1,the virus titer reached 10^(8.67)TCID_(50)/mL.The prepared vaccine showed high safety,of which the protective rate to Cairnamoschata was 100%.Conclusion The procedure for culture of aMPV in bioreactor was optimized,and high virus titer was obtained,which provided a reference for large-scale production of aMPV vaccine.

关 键 词：BHK-21细胞 禽偏肺病毒 悬浮培养 生物反应器 

分 类 号：S855.3[农业科学—临床兽医学]