S/D法及干热法对人凝血因子Ⅷ制品中猪伪狂犬病病毒和猪细小病毒灭活效果的验证

Validation of inactivation effect of porcine pseudorabies virus and porcine parvovirus in human coagulation factor Ⅷ by S/D method and dry heating

目的验证S/D法及干热法对人凝血因子Ⅷ(FⅧ)制品中猪伪狂犬病病毒(pesdorabies virus,PRV)和猪细小病毒(porcine parvovirus,PPV)的灭活效果。方法以PRV和PPV为指示病毒,模拟FⅧ生产工艺中的S/D及干热病毒灭活工艺,采用96孔板微量细胞病变法检测3批FⅧ半成品的残留病毒滴度。结果批号为201203002、2012-04003、201204004 3批FⅧ半成品经(25±1)℃S/D灭活处理15 min后,PRV滴度分别下降≥6.5、≥6.8、≥6.8 lg TCID50/0.1 ml;经(100±2)℃干热灭活30 min后,PPV滴度分别下降4.00、3.92、3.94 lg TCID50/0.1 ml。结论 S/D法及干热法分别对FⅧ制品中PRV和PPV具有良好的灭活效果,本实验为凝血因子类制品的病毒灭活验证工艺的研究奠定了基础。

Objective To verify the inactivation effect of porcine pseudorabies virus（PRV）and porcine parvovirus（PPV）in coagulation factor Ⅷ by S/D method and dry heating.Methods The inactivation procedure during manufacturing of coagulation factor products was simulated using PRV and PPV as indicators.The residual virus titer in three batches of final bulks of FⅧ were determined by micro-CPE on a 96-well microtiter plate.Results The PPV titers in final bulks of FⅧ of Lots 201203002,201204003 and 2012040043 decreased by not less than 6.5,not less than 6.8 and not less than6.8 TCID_（50）/0.1 ml after inactivation by S/D method at（25 ± 1） ℃ for 15 min.However,after dry heating at（100 ±2）℃ for 30 min,the PV titers decreased by 4.00,3.92 and 3.94 Lg TCID_（50）/0.1 ml respectively.Conclusion Both S/D method and dry heating inactivated the PRV and PPV in F Ⅷ effectively,which laid a foundation of further study on verification of virus inactivation procedure of coagulation factor products.