基于huvec细胞表面elam-1表达的抗tnf-α抗体-生物技术通报.pdfVIP

生物技术通报 ·技术与方法· BIOTECHNOLOGY BULLETIN ( ) 2015, 31 12 :70-74 基于 HUVEC 细胞表面 ELAM-1 表达的抗 TNF-α 抗体 活性检测方法 陈坤  徐军  谢灿  周冬梅  杨彬  孙文正 （广东东阳光药业有限公司，东莞 523867） 摘 要 ： 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。 关键词 ： 单克隆抗体 ；HUVEC 细胞；ELAM-1 ；肿瘤坏死因子TNF-α DOI ：10.13560/ki.biotech.bull.1985.2015.12.010 - -α A Method for the Activity Assay of Anti TNF Antibodies Expressed - by ELAM 1 in HUVEC Surface Chen Kun Xu Jun Xie Can Zhou Dongmei Yang Bin Suen Wenzheng （Sunshine Lake Pharma Co. ，Ltd ，Dongguan 523867 ） Abstract: 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