Spectrophotometric Enzyme Assays for High-Throughput Screening
Jean-Louis Reymond*
Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, CH-3012 Bern, Switzerland
Article history:
Received July 18, 2004
Accepted November 22, 2004
Key words:
screening of enzyme activities, TLC-based screening, fluorogenic assay, adrenalin test for enzymes, adrenalin test and copper-calcein assay for amino acids
Summary:
This paper reviews high-throughput screening enzyme assays developed in our laboratory over the last ten years. These enzyme assays were initially developed for the purpose of discovering catalytic antibodies by screening cell culture supernatants, but have proved generally useful for testing enzyme activities. Examples include TLC-based screening using acridone-labeled substrates, fluorogenic assays based on the -elimination of umbelliferone or nitrophenol, and indirect assays such as the back-titration method with adrenaline and the copper-calcein fluorescence assay for amino acids.
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