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

关键词:  IL-17A  BIAcore  生物活性检测  单克隆抗体

Key words:  IL-17A  BIAcore  biological activity test  monoclonal antibody

摘要:

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0.100 86 nmol·L^-1，样品浓度为 2.062 5~ 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-17A单克隆抗体生物活性。

Abstract:

OBJECTIVE To establish two analytical method to determine the biological activity of IL-17A monoclonal antibody. METHODS ①The 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 0.100 86 nmol·L^-1, and the sample concentration was 2.062 5-33 nmol·L^-1. The curve fitting constants R²detected 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.