Nanodroplet profiling of enzymatic activities in a microarray.
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.
Uttamchandani
Mahesh
M
Department of Biological Sciences, National University of Singapore, 3 Science Drive 3, Singapore 117543, Singapore.
Huang
Xuan
X
Chen
Grace Y J
GY
Yao
Shao Q
SQ
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
D010744:Phosphoric Monoester Hydrolases; D010447:Peptide Hydrolases