Miniaturized Stopped-Flow Reaction Microcalorimeter for Biomedical Applications

Principal Investigator Program DirectorLastfirstmiddleBrownRichardKennethPROJECT SUMMARY Calorimetrya universal detection technologymeasures directly the heat released or absorbed by chemical reactionsDespite their potential range of applicationscurrent microcalorimeters exhibit drawbacks that have precluded their use in the most current areas of biomedical researchThese drawbacks derive from their relatively low sensitivityrequirement for large amounts of reactants and low throughputApplications such as enzyme inhibitionscreening for drug candidatescell biologyproteomicsantibiotic profiling and clinical laboratory assays have been inaccessible to calorimetryDuring Phase I of this grantwe developed a miniaturized high sensitivity stopped flow microcalorimeter with a total reaction volume ofLThis instrument has been shown to be able to measure enzyme activity and enzyme inhibition assays with as little enzyme aspicomolesThe miniaturization has been achieved by utilizing novel high density thin film thermoelectric sensors and a stopped flow reaction cell design that eliminates the mechanical stirring that is present in all reaction calorimeters currently in the marketAs the instrument measures directly the heat associated with the enzymatic reaction it does not require any chemical modification or labeling of the substrateas is necessary in standard assaysThe main goal of this Phase II SBIR is to transform the single cell prototype calorimeter successfully completed during Phase I into a high throughputsamples per daycommercial ready multi cell instrument with automated fluid handlingautomated instrument operation and automated data processinganalysis and reportingSince most targets for drug development are enzymes and the search for inhibitors is a major endeavorwe anticipate that the instrument to be developed in this Phase II grant will greatly facilitate and accelerate the drug discovery process and become widely adopted by the pharmaceutical and biotechnology industry Principal Investigator Program DirectorLastfirstmiddleBrownRichardKennethPROJECT NARRATIVE The availability of a multi cell Stopped Flow Reaction Microcalorimeter with a reaction volume ofL per cell will provide a way to perform enzyme activity and inhibition assays for a large number of compoundsCurrent instruments have reaction volumes ofL and are limited to a single cellNew developments in high density thin film thermoelectric chips now enable the development of such miniaturized calorimetric devicesDevelopment of these devices will greatly expand the use of calorimetry in many fundamental and applied biomedical research fieldsThis calorimeter will find use in areas ranging from screening for enzyme inhibitors to clinical laboratories as it allows accurate determination of the concentration of enzymes or substrates in different fluids