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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 conditions for the detection of Fc fragment of intravenous immunoglobulins (IVIG) and to verify the reproducibility of this method. Methods Based on the test of human immunoglobulin Fc fragment activating complement in the three parts of Chinese Pharmacopoeia (2010 edition), the antigen (diphtheria toxoid) titers of sensitized erythrocytes were optimized (25, 50, 100, 150, 200 and 250 Lf / ml), three test conditions of IVIG concentration (5, 10, 20, 30, 40 and 50 mg / ml) and complement titer (15, 50, 75, 100 and 150 CH50 / ml) , Repeated the determination of a group of IVIG products 10 times, calculated coefficient of variation (CV), to verify the repeatability of the method. Results The kinetics of complement-mediated hemolysis showed that the kinetic curves of complement-mediated hemolysis showed that the antigen titer of erythrocytes (diphtheria toxoid) titer of 100 ~ 200 Lf / ml, IVIG concentration of 50 mg / ml and complement titer of ≥ 75 CH50 / ml Typical “S” type, accurate calculation of IVIG Fc fragment activation complement index (IFc). Using optimized test conditions, the determination of one batch of IVIG products was repeated 10 times, the kinetic curves of hemolytic reaction almost coincide as one, and the IFc value of 10 determinations was 8.91%. Conclusion The detection conditions of biological activity of Fc fragment of IVIG are optimized, and the optimized detection method has good repeatability.