终端干热法灭活凝血因子类制品中细小病毒的工艺验证

Validation of Procedure for Inactivation of Parvovirus in Coagulation Factor Products by Terminal Dry Heating

目的验证终端干热法灭活凝血因子类制品中细小病毒的灭活工艺,并对其灭活效果进行评价。方法以猪细小病毒(Porcine parvovirus,PPV)为指示病毒,模拟凝血因子类制品生产工艺中病毒灭活的工艺,采用96孔板微量细胞病变法检测残余病毒滴度,验证终端干热法灭活病毒的效果。结果 3家企业生产的人凝血因子Ⅷ制品经100℃加热30min灭活,病毒滴度分别下降2.01～2.31、≥3.92和3.13～3.50LgTCID50/0.1ml;冻干与加热联合作用,病毒滴度分别下降2.82～3.00、≥4.24和4.00～4.12LgTCID50/0.1ml;5家企业生产的人纤维蛋白原经100℃加热30min灭活,病毒滴度分别下降3.50～3.63、2.56～2.70、3.06～3.57、3.00～3.25和≥3.75LgTCID50/0.1ml;冻干与加热联合作用,病毒滴度分别下降4.00～4.12、3.12～3.46、4.04～4.56、3.50～3.75和≥4.63LgTCID50/0.1ml。结论终端干热法对凝血因子类制品中的细小病毒具有一定的灭活作用,但由于工艺复杂、影响因素较多,不同企业生产的同类制品的病毒灭活效果不尽相同。

Objective To validate the procedure for inactivation of parvovirus in coagulation factor products by terminal dry heating and evaluate the inactivation effect.Methods The inactivation procedure during manufacturing of coagulation factor products was simulated using porcine parvovirus（PPV）as an indicator.The residual virus titer was determined by micro-CPE on a 96-well microtiter plate to validate the inactivation efficacy by terminal dry heating.Results The virus titers in human coagulation Ⅷ manufactured by three manufacturers after inactivation by heating at 100℃ for 30 min decreased by 2.01 ~ 2.31,≥ 3.92 and 3.13 ~ 3.50 LgTCID50 /0.1 ml,while those by lyophilization combined with heating decreased by 2.82 ~ 3.00,≥ 4.24 and 4.00 ~ 4.12 LgTCID50 /0.1 ml,respectively.However,the virus titers in human fibrinogen manufactured by five manufacturers after inactivation by heating at 100℃ for 30 min decreased by 3.50 ~ 3.63,2.56 ~ 2.70,3.06 ~ 3.57,3.00 ~ 3.25 and ≥ 3.75 LgTCID50 /0.1 ml,while those by lyophilization combined with heating decreased by 4.00 ~ 4.12,3.12 ~ 3.46,4.04 ~ 4.56,3.50 ~ 3.75 and ≥ 4.63 LgTCID50 /0.1 ml,respectively.Conclusion Terminal dry heating showed a certain inactivation effect on the PPV in coagulation factor products.However,because of complicated procedure and multiple influencing factors,the virus inactivation efficacies of products of the same kind by various manufacturers were different.