Abstract

Low-volume multiplexed proteolytic activity assay analysis through a pico-injector array

Chen Chia-Hung, Assistant Professor, National University Of Singapore

Secreted active proteases, from families of enzymes such as matrix metalloproteinases (MMPs) and metalloproteinases (ADAMs), participate in diverse pathological processes. To simultaneously measure multiple specific protease activities, a series of parallel enzyme reactions combined with a series of inhibitor analyses for Proteolytic activity matrix analysis (PrAMA) are essential, which reach limitation in sample quantity requirement and the complexity of performing multiple reactions. To address these issues, in this study we developed a pico-injector array to generate 72 different reactions in picoliter-sized droplets by controlling the sequence of combinational injections, which allows simultaneous read-outs of a wide range of multiple enzyme reactions and measurement of inhibitor effects by using small sample volumes (~10?L). Multiple MMPs activates were determined simultaneously by 9 different substrates and 2 inhibitors through injections by a pico-injector array.