基于HUVEC细胞表面ELAM-1表达的抗TNF-α抗体活性检测方法

A Method for the Activity Assay of Anti-TNF-α Antibodies Expressed by ELAM-1 in HUVEC Surface

旨在建立抗TNF-α单克隆抗体生物学活性测定的方法。肿瘤坏死因子TNF-α能刺激人脐静脉内皮细胞HUVEC表达黏附分子ELAM-1,通过抗TNF-α抗体中和TNF-α来抑制这种表达,从而建立该抗体的生物学活性检测方法,并进行验证。结果表明,建立抗体活性检测方法,验证方法的特异性较好、回收率为92.1%-102.1%,方法重复12次,RSD为7.66%。在50%-150%的线性良好,相关系数为0.99。该活性检测方法适于抗体体外活性的检测。

This study is to develop a method to assay the biologic activity of anti-TNF-α（tumor necrosis factor-alpha）monoclonal antibodies. TNF-α stimulates the expression of adhesion molecules E-Selectin（ELAM-1）in human umbilical vein endothelial cells（HUVEC）, whereas anti-TNF-αantibodies suppress the expression of ELAM-1 in HUVEC cells through neutralizing TNF-α. Based on this principle, we developed and validated a method for the bioactivity assay of the antibodies. The result showed that the assay method was established successfully, and the specificity of the method was validated to be feasible. The recovery rate was 92.1%-102.1%, and the RSD was ≦ 7.66% while repeated 12 times. The linear range in 50% to 150% was fine, and the correlation coefficient was 0.99. Conclusively, this method is well suited as an activity assay of antibodies in vitro.