IL-17A单克隆抗体生物学活性检测方法的建立

Establishment of Analytical Method for Biological Activity of IL-17A Monoclonal Antibody

目的建立2种检测IL-17A单克隆抗体的生物学方法。方法(1)利用BIAcore T200系统,基于表面等离子共振技术(surface plasmob resonance,SPR)的分析方法检测IL-17A的生物学活性。(2)采用IL-6释放抑制法测定IL17-A单克隆抗体生物学活性。结果采用SPR法获得的偶联最佳pH值为5.0,动力学常数为0.10086 nmol·L^(-1),样品浓度为2.0625~33 nmol·L^(-1);IL-6释放抑制法检测的曲线拟合常数R2均满足>0.95,相对活性均在80%~125%,多次试验的RSD≤20%,满足重复性验证标准,符合要求。结论2种检测方法的结果均较好,可以有效地反映IL-17A的生物学活性,可以简单、快速用于检测IL^(-1)7A单克隆抗体生物活性。

OBJECTIVE To establish two analytical method to determine the biological activity of IL-17A monoclonal antibody.METHODS①The biological activity of IL-17A was detected by BIAcore T200 analysis method based on surface plasma resonance(SPR).②Determine the biological activity of IL-17A monoclonal antibody by IL-6 release inhibition method.RESULTS The optimum pH value of coupling obtained by SPR method was 5.0,the kinetic constant was 0.10086 nmol·L^(−1),and the sample concentration was 2.0625−33 nmol·L^(−1).The curve fitting constants R2 detected by IL-6 release inhibition method met the requirements of>0.95,the relative activity was 80%−125%,and the RSD of multiple tests was≤20%,which met the repeatability verification standard and met the requirements.CONCLUSION The results of the two detection methods are good,which can effectively reflect the biological activity of IL-17A,and can be used to detect the biological activity of IL-17A monoclonal antibody simply and quickly.