title: Nanodroplet profiling of enzymatic activities in a microarray.
abstract: We describe a generic method for the large-scale functional characterization of enzymes in a microarray. Poly-l-lysine and amine reactive slides were coated with fluorogenic substrates sensitive to proteases and phosphatases. Patterning enzymes on the slides by robotic printing produced spatially addressable, segregated droplets that were simultaneously exposed to the on-chip sensors. Multiple enzymes were profiled using this system that provided fluorescence readouts across temporal and stoichiometric dimensions concurrently on a single microarray substrate. This integrated microarray platform is applicable not only for the functional annotation of proteins, but also for the rapid agonist and antagonist discovery and in performing on-chip kinetics.
authors.affiliation: Department of Biological Sciences, National University of Singapore, 3 Science Drive 3, Singapore 117543, Singapore.
authors.forename: Mahesh
authors.initials: M
authors.lastname: Uttamchandani
authors.forename: Xuan
authors.initials: X
authors.lastname: Huang
authors.forename: Grace Y J
authors.initials: GY
authors.lastname: Chen
authors.forename: Shao Q
authors.initials: SQ
authors.lastname: Yao
mesh_terms: D046228:Microarray Analysis / Q000379:methods*; D049329:Nanostructures / Q000032:analysis*; D036103:Nanotechnology / Q000379:methods; D010447:Peptide Hydrolases / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D010744:Phosphoric Monoester Hydrolases; D010447:Peptide Hydrolases