静注人免疫球蛋白(pH4)Fc段生物学活性检测条件的优化  

Optimization of condition for determination of biological activity of Fc fragment of intravenous immunoglobulin(pH4)

作　　者：何彦林[1] 张璘[1] 张继鹏[1] 张齐明[1] 刘晓[1] 杨晓东[1] 张安山[1] 

机构地区：[1]兰州生物制品研究所有限责任公司血液制剂室,甘肃兰州730046

出　　处：《中国生物制品学杂志》2013年第8期1183-1185,1189,共4页Chinese Journal of Biologicals

摘　　要：目的优化静注人免疫球蛋白(intravenous immunoglobulins,IVIG)Fc段生物学活性检测条件,并验证该方法的重复性。方法基于《中国药典》三部(2010版)人免疫球蛋白Fc段激活补体的试验方法,优化致敏红细胞的抗原(白喉类毒素)效价(25、50、100、150、200和250 Lf/ml)、IVIG浓度(5、10、20、30、40和50 mg/ml)和补体效价(15、50、75、100和150 CH50/ml)3个试验条件;采用优化的试验条件,对1批IVIG制品重复测定10次,计算变异系数(CV),验证该方法的重复性。结果在致敏红细胞的抗原(白喉类毒素)效价100～200 Lf/ml、IVIG浓度50 mg/ml和补体效价≥75 CH50/ml的条件下,补体介导的溶血反应动力学曲线呈典型的"S"型,可准确计算出IVIG Fc段激活补体的指数(IFc)。采用优化的试验条件,对1批IVIG制品重复测定10次,溶血反应动力学曲线几乎重合为一条,10次测定的IFc值的CV=8.91%。结论优化了IVIG Fc段生物学活性检测条件,优化后的检测方法重复性较好。Objective To optimize the condition for determination of biological activity of Fc fragment of intravenous immunoglobulin（IVIG）and verify the reproducibility of the test method.Methods The titer（25,50,100,150,200 and 250 Lf / ml）of diphtheria toxoid for sensitization of red cells,IVIG concentration（5,10,20,30,40 and 50 mg / ml） and complement potency（15,50,75,100 and 150 CH50/ ml）were optimized based on the complement activation test on Fc fragment of human immunoglobulin in Chinese Pharmacopeia（Volume Ⅲ,2010）.One batch of IVIG was determined for 10 times under the optimized condition,of which the CV was calculated,and the reproducibility was verified.Results The optimal diphtheria toxoid titer,IVIG concentration and complement potency were 100 ~ 200 Lf / ml,50 mg / ml and not less than 75 CH50 / ml respectively.Under the optimized condition,the dynamic curve of complement-mediated hemolytic reaction was in a typical S shaped.The index of complement activation by Fc fragment of IVIG（IFc） was calculated accurately.The dynamic curves of complement-mediated hemolytic reaction of one batch of IVIG determined for 10 times under the optimized condition were basically coincident.The CV of 10 test results was 8.91%.Conclusion The condition for determination of biological activity of Fc fragment of IVIG was optimized,and the optimized method showed high reproducibility.

关 键 词：静注人免疫球蛋白 FC段 生物学活性 检测 优化 

分 类 号：R392.33[医药卫生—免疫学]