两种分析冻干人用狂犬病疫苗(Vero细胞)原液及半成品疫苗颗粒含量方法的比较  

Comparison of two methods for content analysis of vaccine particles in freeze-dried human rabies vaccine (Vero cells) bulk and final bulk

作　　者：杨雪 张秋菊 冯子祎 林洪娟 张颖 刘畅 孙宏亮 YANG Xue;ZHANG Qiuju;FENG Ziyi;LIN Hongjuan;ZHANG Ying;LIU Chang;SUN Hongliang(Changchun Institute of Biological Products Co.,Ltd.,Changchun 130062,Jilin Province,China)

机构地区：[1]长春生物制品研究所有限责任公司,吉林长春130062

出　　处：《中国生物制品学杂志》2024年第7期866-872,共7页Chinese Journal of Biologicals

基　　金：吉林省科技发展计划项目(20210401063YY)。

摘　　要：目的 对两种测定冻干人用狂犬病疫苗(Vero细胞)原液及半成品疫苗颗粒含量的方法进行比较分析,为制定狂犬病疫苗制品颗粒含量和纯度标准检测方法提供实验依据。方法 将样品进行疫苗颗粒裂解、BCA定量、体积排阻色谱-高效液相色谱法(size exclusion chromatograghy-high performance liquid chromatography,SEC-HPLC)分析,分别采用BCA-SEC相对面积法和BCA-SEC标准曲线法对冻干人用狂犬病疫苗(Vero细胞)原液及半成品总蛋白浓度进行检测,并对疫苗颗粒含量进行定量分析,对两种方法的检测结果进行对比分析。结果 两种方法测定冻干人用狂犬病疫苗原液疫苗颗粒含量在256~305μg/mL之间,平均值在267~285μg/mL之间,方法间相对标准偏差(relative standard deviation,RSD)在5.8%~10.2%之间,一致性良好。两种方法测定冻干人用狂犬病疫苗半成品疫苗颗粒含量在149~169μg/mL之间,平均值在152~164μg/mL之间,方法间RSD在0.9%~4.7%之间,一致性良好。结论BCA-SEC标准曲线法实测值与预期值偏离较小,更接近样品真实情况,推荐作为企业参考使用。Objective To compare and analyze two methods for determination of vaccine particle content in freeze-dried human rabies vaccine(Vero cells)bulk and final bulk,and to provide experimental basis for establishing standard detection method for particle content and purity of rabies vaccine products.Methods The samples were subjected to vaccine particle cracking,BCA quantification,and size exclusion chromatograghy-high performance liquid chromatography(SEC-HPLC)analysis.BCA-SEC relative area method and BCA-SEC vaccine particle standard curve method were used to detect the total protein concentration of the bulk and final bulk,separately,and the content of vaccine particles was quantitatively analy-zed.Finally,the detection results of the two methods were compared and analyzed.Results The content of vaccine particles in freeze-dried human rabies vaccine bulk was determined to be within 256-305 μg/mL by the two methods,with an average value of 267-285 μg/mL,and the relative standard deviations(RSDs)ranged from 5.8% to 10.2%,with good consistency between two methods.The content of vaccine particles in freeze-dried human rabies vaccine final bulk was determined to be in the range of 149-169 μg/mL by the two methods,with an average value of 152-164 μg/mL,and the two methods showed the RSDs between 0.9%-4.7% with good consistency.Conclusion The measured value by BCA-SEC standard curve method deviates less from the expected value and is closer to the actual situation of samples,so it is recommended to use this method as a reference for enterprises.

关 键 词：狂犬病疫苗 疫苗原液 半成品 病毒颗粒 批间一致性 含量分析 

分 类 号：Q816[生物学—生物工程]