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A Droplet Microarray for Homogeneous Enzymatic Assay

M. Balakirev, L. Mugherli and F. Chatelain
CEA-Grenoble, FR

microarray, droplets, screening, proteases, fluorescence

This work describes a droplet microarray for monitoring of enzymatic activity on a glass slide. These array are composed of features defined and separated by differential surface tension (surface tension array). A photolithographic method was used to create a series of circular hydrophilic spots delimited by a perfluorosilanated surface. The perfluorinated region had water contact angle on the order of 105°, whereas inside the spots this angle was < 75°. The difference in surface tension “holds” water within the hydrophilic spots thus resulting in droplet microarray. We show that each droplet can be used as a micro-reservoir for conducting of enzymatic reactions. As a proof of concept a proteolytic cleavage of a fluorogenic substrate was carried out within hundreds of confined reservoirs created on a glass slide. Using piezoelectric deposition of substrate and protease numerous reaction conditions could be tested in parallel. This format may be a valuable tool for high-throughput screening, and enzymatic profiling of novel proteins.

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