重组人凝血因子Ⅷ病毒灭活/去除工艺验证  

Verification of viral inactivation/removal process for recombinant human coagulation factor Ⅷ

作　　者：黄娟 李德款 武志强 宋春雷 李成 容新宗 寇鹏 雷韬 HUANG Juan;LI De-kuan;WU Zhi-qiang;SONG Chun-lei;LI Cheng;RONG Xin-zong;KOU Peng;LEI Tao(Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education,College of Life Sciences,Sichuan University,Chengdu 610065,Sichuan Province,China)

机构地区：[1]四川大学生命科学学院生物资源与生态环境教育部重点实验室,四川成都610065 [2]成都蓉生药业有限责任公司科研开发部,四川成都610041 [3]成都生物制品研究所有限责任公司,四川成都610023

出　　处：《中国生物制品学杂志》2019年第12期1411-1414,共4页Chinese Journal of Biologicals

摘　　要：目的 验证重组人凝血因子Ⅷ(recombinant human coagulationⅧ,rhFⅧ)病毒灭活/去除工艺。方法 采用S/D法作为rhFⅧ病毒灭活工艺,纳米过滤(20 nm孔径)作为rhFⅧ病毒去除工艺,分析病毒灭活/去除两步工艺对rhFⅧ活性的影响;分别以伪狂犬病毒(pseudorabies virus,PRV)、水疱性口炎病毒(vesicular stomatitis virus,VSV)、小鼠白血病病毒(xenotropic murine leukemia virus,X-MuLV)及小鼠细小病毒(murine minute virus,MMV)作为指示病毒,对上述工艺进行病毒灭活/去除效果验证。结果 S/D灭活工艺中,3批制品的活性收率分别为96. 2%、100. 4%和103. 2%;(24±2)℃处理180 min后,指示病毒PRV、VSV和X-MuLV滴度降低量(Log10)虽然均<4,但盲传3代,仍未检出病毒。纳米过滤工艺中,3批制品蛋白回收率均大于98%,活性回收率均大于90%,纳米过滤120 min,MMV滴度降低量(Log10)> 4。结论 建立的rhFⅧ病毒灭活/去除工艺,可有效灭活/去除病毒,保证制品的质量和安全。Objective To verify the viral inactivation/removal process for recombinant human coagulation factor Ⅷ(rhF Ⅷ).Methods The solvent/detergen(S/D)method was used as virus inactivation process,while the nanofiltration(20 nm pore size)as virus removal process for rhF Ⅷ to analyze the effect of the two-step virus inactivation/removal process on the biological activity of the product.Pseudorabies virus(PRV),vesicular stomatitis virus(VSV),xenotropic murine leukemia virus(X-MuLV)and murine minute virus(MMV)were used as indicators to verify the inactivation/removal effect of the above process,respectively.Results In the S/D inactivation process,the recovery rates of activities of the three batches of the products were 96.2%,100.4%and 103.2%,respectively.After treatment at(24±2)℃for 180 min,the titers of PRV,VSV and X-MuLV decreased by less than 4 Log10,while no virus was detected after three blind passages.However,in the nanofiltration process,all the protein recoveries of three batches were more than 98%,while the specific activities were more than 90%.After nanofiltration for 120 min,the titer of MMV decreased by more than 4 Log10.Conclusion The developed processes were effective in inactivation/removal of viruses in rhF Ⅷ to ensure the quality and safety of products.

关 键 词：重组人凝血因子Ⅷ S/D法 纳米过滤 灭活 去除 

分 类 号：R392-33[医药卫生—免疫学]