Development of a p38未 mitogen activated protein kinase ELISA assay for the quantitative determination of inhibitor activity

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• 作者：Má ; rcia Goettert^a ; Nouran Shaalan^b ; Ralph Graeser^c ; Stefan A. Laufer^a ; ^{stefan.laufer@uni-tuebingen.de}
• 关键词：MAPK ; mitogen-activated protein kinase ; RASFs ; rheumatoid arthritis synovial fibroblast ; ATF-2 ; activating transcription factor 2 ; ELISA ; enzyme-linked immunosorbent assay ; MMPs ; matrix metalloproteases ; RA ; rheumatoid arthritis ; TBS ; tris-buffered saline
• 刊名：Journal of Pharmaceutical and Biomedical Analysis
• 出版年：2012
• 期刊代码：65_07317085
• 类别：ch
• 出版时间：July, 2012
• 卷：66
• 期：Complete
• 页码：349-351
• 文件大小：185 K

摘要

The p38 mitogen activated protein kinase (MAPK) has emerged as a target for treating inflammatory diseases, like rheumatoid arthritis (RA). Expression of p38未 is induced in rheumatoid arthritis synovial fibroblasts (RASFs) by a cytokine-independent pathway substantially different from other MAPK pathways. To identify inhibitors of p38未 MAPK, we developed a direct ELISA assay based on a previously described p38伪 assay for monitoring the phosphorylation of ATF-2. This work presents a straightforward assay for evaluating the potency of small-molecule inhibitors. To validate the assay under optimized conditions, we used reference compounds and achieved results comparable to published data.