Enzyme kinetics studies to guide mathematical modeling of microdialysis sampling to predict in situ biochemistry

by Klucher, Justin; Stenken, Julie Ann

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This study aims to build a kinetic understanding of the enzyme porcine pancreatic elastase, a human neutrophil elastase mimic, in order to apply microdialysis sampling methodol. to in vivo biochem. monitoring of the following reaction: Succinyl-(ala)3-p-nitroanilide + Elastase ? p-nitroaniline + Elastase. A previously available microdialysis mass transport model is being used to model the in situ biochem. of this reaction. Microdialysis parameters of delivery of substrate and recovery of product under various combinations of enzyme and substrate concns. are being evaluated as a means to predict enzymic activity external to the microdialysis probe. Michaelis-Menten kinetic parameters for porcine pancreatic elastase are incorporated into the math. model for ref. Then using a microdialysis mass transport model, this kinetic information will be used to predict recovered product concns. This data will then be used to attempt to monitor in situ systems with a reliable level of activity prediction.