尿胰蛋白酶抑制剂生产中病毒灭活工艺的效果观察

Investigation of virus inactivation methods used in the production of urinary trypsin inhibitor(UTI)

目的验证尿胰蛋白酶抑制剂(UTI)生产工艺中采用的60℃水浴10 h和乙醇处理3 h的病毒灭活效果。方法将Sindbis病毒、伪狂犬病毒(PRV)和脊髓灰质炎病毒(PV1)3种指示病毒分别加入不同的UTI原料样品中,进行60℃水浴10 h和乙醇处理3 h,处理不同时间后分别取样,用微量细胞病变法检测不同时间段样品中的病毒残留滴度。结果60℃水浴10 h可灭活(6.503±0.102)LgTCID50/mL的Sindbis病毒,灭活(6.42±0.158)LgTCID50/mL的PRV以及(6.587±0.061)LgTCID50/mL的PV1。乙醇处理3 h后可灭活(5.88±0.204)LgTCID50/mL的Sindbis病毒,灭活(6.378±0.268)LgTCID50/mL的PRV以及(5.963±0.118)LgTCID50/mL的PV1。经两种方法处理后的样品加入细胞经3代盲传均未见细胞病变(CPE)出现。结论UTI生产工艺中采用的60℃水浴10 h和乙醇处理3 h均可灭活所验证的3种指示病毒,灭活效果>6.0 LgTCID50/mL。

Purpose To test the virus inactivation effect of water bath method at 60 ℃ for 10 hours and alcohol treatment for 3 hours which was used in the production of urinary trypsin inhibitor（UTI）. Methods Sindbis virus, Pseudorabies virus（PRV） and poliovirusl （PV1） were used as indicated viruses in this test. After being added separately into the UTI raw material in 10% proportion,the viruses were treated with water bath at 60 ℃ for 10 hours and alcohol for 3 hours and then the samples of UTI were taken to inoculate the cell line for assay of cytopathic effect. Results The water bath at 60 ℃ for 10 hours could inactive Sindbis, PRV and PV1 in more than （ 6. 503 ± 0.102 ） LgTCID50, （ 6.42 ± 0.158 ） LgTCID50 and （6.587 ± 0.061 ） LgTCID50 respectively,and alcohol treatment for 3 hours could inactive Sindbis,PRV and PV1 in more than（5.88 ±0.204）LgT- CID50, （6. 378 ±0. 268） LgTCIDso and （5. 963 ±0.118 ） LgTCID50 respectively. No cytopathie effect was found in the cell line which was inoculated with treated samples after blind passage for three generations. Conclusion The water bath method at 60 ℃ for 10 hours and alcohol treatment for 3 hours which were used in the production of UTI had good effects on virus inactivation and the inactivation efficiency on Sindbis, PRV and PV1 was more than 6 LgTCID50/mL.