人纤维蛋白原干热病毒灭活效果及灭活后稳定性的研究  

Study on inactivation effect of human fibrinogen drying heat virus and its stability after inactivation

作　　者：刘环 毕晓峰 张良 宋天旭 江豪琨 江砚芳 LIU Huan;BI Xiao-feng;ZHANG Liang;SONG Tian-xu;JIANG Hao-kun;JIANG Yan-fang(Department of Blood Preparation,Sinopharm Shanghai Plasma-derived Biotherapies Co.,Ltd.,Shanghai 200051,China)

机构地区：[1]国药集团上海血液制品有限公司血液制剂室,上海200051

出　　处：《生物医学工程与临床》2025年第2期155-159,共5页Biomedical Engineering and Clinical Medicine

摘　　要：目的评价人纤维蛋白原(Fg)干热病毒灭活效果及灭活后成品稳定性的研究。方法首先选择连续3批次制备的Fg样品,按样品与病毒9∶1的体积比例分别加入猪细小病毒(PPV)和脑心肌炎病毒(EMCV),冻干后于80℃~87℃条件下干热灭活72 h,使用96孔板微量细胞病变法测定病毒滴度降低量,考察Fg干热灭活效果;然后对干热前/后样品进行蛋白含量、Fg含量、纯度和凝固活力指标的检测并进行比较,以评价其质量变化;最后对干热灭活后的Fg成品进行加速稳定性和长期稳定性研究,考察各项质量指标(如外观、复溶时间、可见异物、水分含量、渗透压摩尔浓度、纯度和凝固活力等)与0个月相比的变化情况,并分析各批次间指标的差异性。其中,加速实验在第0、3和6个月月末抽样检测,长期实验在第0、3、6、9、12、18、24、36和42个月月末抽样检测。结果冻干工艺结合80℃~87℃干热灭活72 h后PPV和EMCV滴度降低量均大于4.0 LgTCID50/0.1 mL,干热灭活前/后Fg质量差异无统计学意义(P>0.05);加速实验和长期实验结果显示各项质量指标的检测结果与0个月相比,其中外观和可见异物指标无明显变化,其他指标均有所变化,但均满足《中华人民共和国药典》2020版(三部)规定,且各批次间指标差异无统计学意义(P>0.05)。结论冻干人纤维蛋白原采用80℃~87℃干热灭活72 h工艺能有效去除PPV和EMCV,对产品质量影响较小,且最终成品的安全性和稳定性较好。Objective To evaluate the effect of drying heat virus inactivation of human fibrinogen(Fg)and stability of the product after inactivation.Methods Firstly,three consecutive batches of Fg samples were prepared,the porcine parvovirus(PPV)and encephalomyocarditis virus(EMCV)were added by volume ratio of sample to virus 9∶1,respectively,and inactivated at 80℃-87℃for 72 hours after lyophilization.The reduction of virus titer was determined by using the 96-well microplate cytopathic effect method.Then,the protein content,Fg content,purity and coagulation activity of samples before and after drying heat were detected and compared to evaluate the quality changes.Finally,the accelerated stability and long-term stability of Fg finished products after drying heat inactivation were studied.The changes of various quality indicators(such as appearance,re-dissolution time,visible foreign matter,moisture content,osmotic pressure molar concentration,purity and coagulation activity)compared with 0 month were investigated,and the differences of indicators between batches were analyzed.At the end of 0,3 and 6 months,samples were tested for accelerated test.At the end of 0,3,6,9,12,18,24,36 and 42 months,samples were tested for long-term test.Results The titers of PPV and EMCV decreased more than 4.0 LgTCID50/0.1 mL after drying heat inactivation at 80°C-87°C for 72 hours,and there was no significant difference in Fg quality before and after drying heat inactivation(P>0.05).The results of accelerated and long-term experiments results showed that there was no significant change in appearance and visible foreign body indexes compared with 0 month,and other indexes changed but met the Pharmacopoeia of the People’s Republic of China 2020 Edition(Part III),and there was no significant difference in indexes between batches(P>0.05).Conclusion It is demonstrated that the drying heat inactivation process could effectively remove PPV and EMCV from lyophilized Fg,which has little effect on product quality,and the finished product has g

关 键 词：人纤维蛋白原 干热灭活 质量 稳定性 差异性 

分 类 号：TQ464.7[化学工程—制药化工]