超滤法去除抗乙肝转移因子中病毒工艺的验证  

Validation of the virus removal by ultrafiltration

作　　者：杨国成[1] 李汝霖[1] 夏腊菊[1] 汤华东 欧阳怡 

机构地区：[1]武汉大学医学院,湖北武汉430071 [2]武汉海特生物制药股份有限公司,湖北武汉430056

出　　处：《中国生化药物杂志》2004年第6期334-335,338,共3页Chinese Journal of Biochemical Pharmaceutics

摘　　要：目的对超滤法去除病毒工艺进行验证。方法以猪细小病毒 (PPV)为指示病毒 ,观察用截留相对分子质量 (Mr)为 5 0 0 0的超滤膜在进压 0 .14MPa、回流压 0 .0 7MPa和进压 0 .2MPa、回流压 0 .1MPa条件下去除抗乙肝转移因子原液中病毒的效果 ,并以淋巴细胞提取液为供试品 ,在上述条件下超滤 ,用高效液相色谱法测定滤过液中各组分的Mr,以检查膜的完整性。结果PPV滴度为 6 .4logTCID50 /ml时 ,滤过液中未检测出PPV ,经盲传三代 ,亦未发现特异性细胞病变 ;滤过液中各组分Mr 均小于 10 0 0 0 ,表明超滤膜完整。PurposeTo validate the virus removal by ultrafiltration.MethodsAnti-HBV transfer factor solution spiked with the porcine parvovirus (PPV) is ultrafiltered with a 5 000 molecular weight cut-off membrane under different work pressures for 1 or 3 cycles. The cell extracts prepared from lymph gland with molecular weight within 100 000 are applied to validate the integrity of the membrane by determining the molecular weight of all components in the filtrate by HPLC.ResultsThe initial titre of PPV spiked into Anti-HBV transfer factor solution is 6.4 LogTCID 50/ml. The residual virus was undetectable in the ultrafiltrate, whether the ultrafiltration procedure was carried out under a feed pressure of 0.14 MPa and under a retentate pressure of 0.07 MPa, one or three cycles; or under a feed pressure of 0.14 MPa and under a retentate pressure of 0.07 MPa, one or three cycles. Cytopathic effects were not observed in three blind passages of the ultrafiltrate samples. It was showed that the membrane is integrated for the molecular weight of all components in the ultrafiltrate and is lower than 10 000.ConclusionThe method of PPV removal by ultrafiltration is effective.

关 键 词：病毒去除法 超滤法 猪细小病毒 高效液相色谱 

分 类 号：TQ460.6[化学工程—制药化工] R392-33[医药卫生—免疫学]