Abstract 4541: Highly sensitive kinase assays combining Carna Biosciences QSS Assist ELISA reagents with the Meso Scale Discovery MULTI-ARRAY platform.

Abstract

Abstract We developed a sensitive kinase asay using a QSS Assist ELISA kinase assay kit from Carna Biosciences, Inc. and the detection method from Meso Scale Discovery. The QSS Assist ELISA kit is optimized for screening of compounds. Comparing to most other kinase assay kits, QSS Assist ELISA kits are available for both peptide and protein substrates and several ELISA kits developed by Carna Biosciences employ a naturally occurring protein substrate, e.g. MAP kinases for MAPK kinases. Detection of kinase activity is achieved with a primary anti-phosphoserine/threonine or tyrosine antibody and a secondary anti-Ig-antibody. Commonly, such secondary antibodies are coupled to horseradish peroxidase (HRP), which allows for signal detection by adding a color reagent, i.e. an HRP substrate which can be detected upon reaction by measuring optical density. Here we applied an MSD SULFO-TAG labeled secondary antibody. These antibodies emit light upon electrochemical stimulation initiated at the electrode surfaces of MULTI-ARRAY microplates, and thus detect the primary antibody with ultimate sensitivity without direct labeling it. We have sucessfully miniaturized the highly sensitive assay and this new assay technology has been validated with published kinase inhibitors from AstraZeneca and Tocris Bioscience. Citation Format: Carsten Jacobi, Alain Schilb, Yusuke Kawase, Yasuyuki Kirii, Brigitte Boldyreff. Highly sensitive kinase assays combining Carna Biosciences QSS Assist ELISA reagents with the Meso Scale Discovery MULTI-ARRAY platform. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4541. doi:10.1158/1538-7445.AM2013-4541