ENHANCED ELISA PERFORMANCE USING ACOUSTIC MICROSTREAMING

SOLID-PHASE, ENZYME-LINKED IMMUNOSORBENT ASSAYS (ELISA'S), SUCH AS THE ELISA AIDS SCREENING TEST, DEPEND ON THE DIFFUSION OF MACROMOLECULES AND ORGANIC SUBSTRATES ACROSS A QUIESCENT LIQUID BOUNDARY LAYER PRIOR TO ADSORPTION TO THE SOLID PHASE OR REACTIVITY WITH SOLID-PHASE-BOUND MACROMOLECULES. TIME-DEPENDENT DIFFUSION OFTEN RESULTS IN LENGTHY INCUBATION TIMES, OR ELEVATED CONCENTRATIONS OF EXPENSIVE OR RARE REAGENTS TO REDUCE INCUBATION TIME. STIRRING BY VIGOROUS MICROPLATE AGITATION IS ONLY MARGINALLYEFFECTIVE IN IMPROVING DIFFUSION, CANNOT BE DONE WHILE THE ENZYME-SUBSTRATE REACTION IS READ IN FAST KINETIC APPLICATIONS, AND PRESENTS POTENTIAL BIOHAZARD AND INSTRUMENT MALFUNCTION PROBLEMS. THE GOAL OF THIS RESEARCH IS TO DETERMINE THE EFFICACY OF ACOUSTIC MICROSTREAMING FOR IMPROVING ELISA PERFORMANCE. THE AIMS ARE: (1) TO USE AN EXISTING ACOUSTIC MICROSTREAMINGPROTOTYPE DEVICE TO DETERMINE OPTIMUM CONDITIONS TO ENHANCE BINDING OF ELISA IMMUNOREACTANTS TO MICROPLATE WELLS, BOTH AT ROOM AND ELEVATED TEMPERATURES; AND (2) TO DETERMINE THE VALUE OF ACOUSTICALLY ENHANCED ELISA PERFORMANCE AS MEASUREDBY INCREASED SENSITIVITY, DECREASED INCUBATION TIME, AND DECREASED REAGENT EXPENDITURE. THE LONG-TERM OBJECTIVE (PHASE II) IS TO DEVELOP AN ACOUSTICMICROPLATE ACOUSTIC MICROSTREAMING DEVICE WITH ELEVATED TEMPERATURE INCUBATOR.