Journal
ANALYTICAL BIOCHEMISTRY
Volume 281, Issue 1, Pages 62-67Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1006/abio.2000.4501
Keywords
enzyme kinetics; mathematics; data analysis; statistics; tight binding; inhibition constant; high-throughput screening; mast-cell tryptase
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Determination of tight-binding inhibition constants by nonlinear least-squares regression requires sufficiently good initial estimates of the best-fit values. Normally an initial estimate of the inhibition constant must be provided by the investigator. This paper describes an automatic procedure for the estimation of tight-binding inhibition constants directly from dose-response data. Because the procedure does not require human intervention, it was incorporated into an algorithm for high-throughput screening of enzyme inhibitors. A suitable computer program is available electronically (http://www.biokin.com). Representative experimental data are shown for the inhibition of human mast-cell tryptase. (C) 2000 Academic Press.
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