基于单克隆抗体的酶联免疫吸附测定法的开发，用于检测gp130与IL-6/IL-6R复合物的结合及其竞争性抑制作用。

Development of a monoclonal antibody-based enzyme-linked immunoabsorbent assay for the binding of gp130 to the IL-6/IL-6R complex and its competitive inhibition.

作者信息

Scheller Jürgen, Kovaleva Marina, Rabe Björn, Eichler Jutta, Kallen Karl-Josef, Rose-John Stefan

机构信息

Institut für Biochemie, Christian-Albrechts Universität zu Kiel, Olshausenstr.40, D-24098 Kiel, Germany.

出版信息

J Immunol Methods. 2004 Aug;291(1-2):93-100. doi: 10.1016/j.jim.2004.05.002.

DOI:10.1016/j.jim.2004.05.002

PMID:15345308

Abstract

The proinflammatory cytokine IL-6 binds to the membrane bound or soluble IL-6 receptor (IL-6R) and activates an intracellular signaling cascade after complex formation with two gp130 molecules. These mediate general homeostasis and orchestrates the immune response during disease. Trans-signalling via the soluble IL-6R has importance for the development and maintenance of human diseases like Crohn's disease, peritonitis and rheumatoid arthritis. We have developed an enzyme-linked immunoabsorbent assay (ELISA) that detects the binding of gp130 to the IL-6/sIL-6R complex. Competitive binding of sgp130-Fc, viral IL-6, and the inhibitory drug Suramin to gp130 has been demonstrated. The assay can be used for high-throughput screening of peptide and chemical compound libraries for the identification of new agonists and antagonists of the binding between gp130 and IL-6/sIL-6R.

摘要

促炎细胞因子白细胞介素-6（IL-6）与膜结合型或可溶性白细胞介素-6受体（IL-6R）结合，并在与两个gp130分子形成复合物后激活细胞内信号级联反应。这些分子介导总体内环境稳定，并在疾病期间协调免疫反应。通过可溶性IL-6R的转信号传导对于克罗恩病、腹膜炎和类风湿性关节炎等人类疾病的发展和维持具有重要意义。我们开发了一种酶联免疫吸附测定法（ELISA），可检测gp130与IL-6/可溶性IL-6R复合物的结合。已证明可溶性gp130-Fc、病毒IL-6和抑制性药物苏拉明对gp130具有竞争性结合。该测定法可用于对肽和化合物文库进行高通量筛选，以鉴定gp130与IL-6/可溶性IL-6R之间结合的新激动剂和拮抗剂。