摘要
目的分析在低温乙醇蛋白分离法基础上引入阴离子交换层析(anion exchange chromatography,AEX)纯化工艺处理,对静注人免疫球蛋白(pH 4)制品质量的影响。方法检测并对比分析低温乙醇蛋白分离法及联合AEX纯化工艺制备的静注人免疫球蛋白(pH 4)制品分子量大小分布、抗补体活性(anticomplement activity,ACA)、抗-HBs效价、白喉抗体效价、IgA含量、抗体Fc段生物学活性、IgG亚型分布、N-糖基化类型、蛋白质二级结构和三级结构的异同。结果在低温乙醇蛋白分离法基础上,引入AEX纯化工艺处理,可大幅度降低静注人免疫球蛋白(pH 4)制品中IgA含量;有利于去除制品中三级结构发生改变的蛋白质;未对制品其他质量指标产生显著影响。结论在低温乙醇蛋白分离法基础上,引入AEX纯化工艺可提高静注人免疫球蛋白(pH 4)制品质量。
Objective To analyze the effect of anion exchange chromatography(AEX)on quality of human immunoglobulin(pH 4)for intravenous injection products based on low temperature ethanol protein separation.Methods Human immunoglobulin(pH 4)for intravenous injection prepared by low-temperature ethanol protein separation method alone and in combination with AEX were determined for molecular weight distribution,anti-complement activity(ACA),anti-HBs titer,diphtheria antibody titer,IgA content,antibody Fc fragment activity,IgG subtype distribution,N-glycosylation type,secondary and tertiary structures of protein,and the results were compared and analyzed.Results Based on the low temperature ethanol protein separation method,the introduction of AEX decreased the IgA content in human immunoglobulin(p H 4)for intravenous injection products significantly,while was beneficial to removal of proteins with tertiary structure damaged,which showed no significant impact on other quality indicators of the product.Conclusion Based on the low temperature ethanol protein separation method,the introduction of AEX improved the quality of human immunoglobulin(pH 4)for intravenous injection products.
作者
张学成
马小伟
安文琪
梁雪爽
罗坚
王承勖
吴静娟
李光飞
ZHANG Xue-cheng;MA Xiao-wei;AN Wen-qi;LIANG Xue-shuang;LUO Jian;WANG Cheng-xu;WU Jing-juan;LI Guang-fei(Hualan Biological Engineering Inc.,Xinxiang 453003,Henan Province,China)
出处
《中国生物制品学杂志》
CAS
CSCD
2019年第9期1036-1042,共7页
Chinese Journal of Biologicals
关键词
静注人免疫球蛋白
阴离子交换层析
IGA
Human immunoglobulin for intravenous injection
Anion exchange chromatography(AEX)
IgA
