= Harold B. Brooks
{c}
<Harold B. Brooks> coauthored practical guidance for designing <enzyme assays> used to test many candidate compounds. His 2012 chapter explains how <enzyme kinetics>, the choice of <reaction substrate> and controls determine whether an assay can identify <reversible enzyme inhibition>. Its emphasis on initial rates connects an instrument's signal to a defined biochemical reaction rather than treating an endpoint measurement as self-explanatory.<Basics of Enzymatic Assays for HTS by Harold B. Brooks and others>[Abstract, Introduction and Concept]{ref}
For example, imagine an <enzyme assay> that starts with $100$ micromolar <reaction substrate>. Measuring after only $5$ micromolar has been consumed leaves its concentration approximately unchanged; measuring after $80$ micromolar has been consumed does not. The chapter recommends establishing an early linear interval with little substrate depletion, checking several <enzyme> concentrations and measuring background signals with appropriate missing-component controls. A later plateau can reflect exhausted <reaction substrate> rather than loss of <enzyme> activity. This is why an <enzyme inhibition initial-rate assay> needs a time course before it can turn a smaller signal into evidence about inhibition.<Basics of Enzymatic Assays for HTS by Harold B. Brooks and others>[What is initial velocity?; Measuring initial velocity of an enzyme reaction; reaction progress curves]{ref}
Back to article page
