应用酶联免疫法测定人免疫球蛋白类制品中IgA含量

目的：建立人免疫球蛋白类制品中免疫球蛋白A（IgA）含量测定的酶联免疫（ELISA）方法。方法：使用筛选出的ELISA检测试剂盒和IgA国际标准品，建立IgA含量测定的ELISA方法并进行方法学验证，使用验证的方法对来自不同企业的22批静注人免疫球蛋白（pH 4）中IgA含量进行测定。结果：建立的方法在1.562 5×10^-3~2.5×10^-4 IU·mL^-1范围内线性良好（r=0.996 3）；与IgG1类单抗药物未显示交叉反应；定量限为2.167×10^-4 IU；含不同辅料的2种处方平均加标回收率分别为102.2%和98.5%；重复性和中间精密度RSD均小于10%；高、中、低浓度样品使用不同批号试剂盒测定结果RSD均小于10%。由于生产工艺不同，各家企业所生产的静注人免疫球蛋白（pH 4）中IgA含量各不相同，经纯化的产品IgA含量更低。结论：该检测方法具有快速、灵敏、操作简便等特点，可用于人免疫球蛋白类制品的质量控制。

Objective:To establish an enzyme-linked immunosorbent assay(ELISA)method for quantification of immunoglobulin A(IgA)in human immunoglobulin products.Methods:A screened ELISA kit and the WHO International Standard(Immunoglobulins G,A and M,Human Serum)were used to set up the ELISA method for quantification.22 batches of human immunoglobulin for intravenous injection(pH 4)from different manufacturers were analyzed using the verified method.Results:The established method demonstrated good linearity in 1.562 5&#215;10^-3-2.5&#215;10^-4 IU&#183;mL^-1(r=0.996 3)without cross-reaction with other immunoglobulins G1 products.The limit of quantitation was 2.167&#215;10^-4 IU.The average recoveries for two formulations with different excipients were 102.2% and 98.5%,respectively.The RSD of repeatability and intermediate precision were both lower than 10%.The ELISA kit also showed good lot-to-lot uniformity with RSD<10% in analyzing different batches of samples at different concentrations.In addition,due to different manufacturing processes,the contents of IgA were different in 22 batches of samples and lower in purified samples.Conclusion:The method is quick,sensitive and convenient,and it can be applied to quality control of human immunoglobulin products.