人凝血酶原复合物短波紫外灭活细小病毒验证及对产品质量的影响  

Validation of Inactivation of Parvovirus in Human Prothrombin Complex Concentrate by Short-wave Ultraviolet C and Impaction on Product Quality

作　　者：王敏[1] 岳广智[1] 杨立宏[1] 贾丽丽[1] 杨靖清[1] 周倩[1] 郝杰[1] 王菁舟[1] 侯继锋[1] 

机构地区：[1]中国食品药品检定研究院,北京100050

出　　处：《中国生物制品学杂志》2011年第5期600-603,612,共5页Chinese Journal of Biologicals

基　　金：国家科技支撑计划项目(2008BAI54B08)

摘　　要：目的应用短波UVC对人凝血酶原复合物(Prothrombin complex concentrate,PCC)进行灭活处理,验证病毒灭活效果及对产品质量的影响。方法以猪细小病毒(Porcine parvovirus,PPV)作为指示病毒,采用短波UVC对PCC样品进行灭活,紫外辐射剂量分别为200、250和300 J/m2,微量细胞培养法检测病毒滴度,对未出现病变的样品盲传3代,验证灭活效果;应用二维电泳(2-DE)、高效液相色谱技术(HPLC)及活性检测方法对PCC样品的结构和功能及活性进行评价。结果短波UVC法可使PPV滴度降低4.0 log以上,所有样品在盲传第3代时均检测到细胞病变。在无保护剂的情况下,凝血因子活性回收率大于70%;HPLC分析结果证实,UVC照射后,凝血因子蛋白聚合体含量无明显改变,与照射前比较,蛋白的色谱行为基本一致;2-DE检测显示,样品照射前后,图谱蛋白斑点数量基本相等,其中300 J/m2 UVC处理组有2个蛋白点群出现位置和灰度值的变化。结论短波UVC对无胞膜病毒灭活效果较好,且该灭活方法对PCC制品蛋白活性影响较小。Objective To inactivate the parvovirus in human prothrombin complex concentrate（PCC） by using short-wave ultraviolet（UVC）,validate the inactivation effect as well as the impaction on product quality.Methods The porcine parvovirus（PPV） as an indicator virus in PPC was inactivated with short-wave UVC at dosages of 200,250 and 300 J / m2 respectively.The virus titer was determined by micro-cell culture method.The samples showing no CPE were subjected to 3 blind passages and validated for inactivation effect.The PPC samples were evaluated for structure and function by 2-DE and HPLC,and determined for activity.Results The titer of PPV inactivated with short-wave UVC decreased by more than 4.0 log.CPEs were observed in all the samples after 3 blind passages.The recovery rate of coagulation factor activity in absence of stabilizer was more than 70%.HPLC proved that the coagulation factor protein polymer content and chromatographic behavior of the samples after UVC irradiation showed no significant change as compared with that before irradiation.2-DE showed that the number of protein spots after irradiation was basically consistent with that before irradiation.However,changes were observed in positions and gray values of two protein spot groups in the samples treated with 300 J / m2 UVC.Conclusion Short-wave UVC showed good effectiveness in inactivation of non-enveloped viruses,with little impaction on protein activity of PCC.

关 键 词：UVC 细小病毒 猪 病毒灭活 人凝血酶原复合物 生物学活性 

分 类 号：R373.9[医药卫生—病原生物学] R392-33[医药卫生—基础医学]