重组人CD11α单克隆抗体生物学活性测定方法的优化

Improvement Method for Detection of Recombinant Humanized Monoclonal Antibody CD11α Biological Activity

用竞争性荧光免疫抑制细胞实验法对重组人CD11α单克隆抗体注射剂进行生物学活性测定,对LFA1转染的293细胞的培养、抗体包被时间及活性测定过程中样品的制备等条件进行了优化,建立了规范可行的操作方法。3批样品的生物学活性分别为:1.03×104U/mg蛋白,0.98×104U/mg蛋白及0.97×104U/mg蛋白,5次测定的RSD分别为4.554、5.039及1.443。

To improve the current method which has been used for detection of recombinant humanized monoclonal antibody CD11α biological activity, competitive fluorescent immunity inhibition method has been used in this study. This method is based on 293 cell transfected with CD11α subunit of LFA1 which can bind to ICAM-IgG and MAb CD11α inhibits the binding of LFA-1 to ICAM-1. The biological activities of 3 batches of imported recombinant humanized monoclonal antibodies CD11α provided by manufacturer have been measured. The biological activities of these 3 batches samples are 1.03 × 10^4 U/mg, 0.98 × 10^4 U/mg and 0.97 × 10^4 U/mg respectively, RSD（5 times of measurement） are 4. 554, 5. 039, 1. 443 respectively. The modified method by changing the cell culture condition, coating time of antibody as well as the solution condition for sample preparation can be used as standard method for detection of recombinant humanized monoclonal antibody CD11α biological activity.