ROUTINE DETERMINATION OF ENZYME-KINETICS USING PLATE READER

A method is described for the routine determination of kinetic parameters from colorimetric enzyme reactions using a 96-well microtiter plate reader commonly used in immunoassay. A program in Turbo Pascal have been developed for the analysis of results. The system performs the description of plate format, the definition of conditions of assays, the measurement of enzyme reactions, the collection of data, analysis, visualization of results, and modification of data. Measurement of K(m) and V(max) is made with different methods, including iterative determination, Lineweaver-Burk transformation, and Hofster transformation, to give the final values of kinetic parameters with their associated standard errors. This approach should be applicable to routine determination of kinetic parameters.