title: Array-on-Array Strategy For Activity-Based Enzyme Profiling.
abstract: We describe a novel array on array strategy intended to enhance the throughput of enzymatic activity screening using microarrays. This strategy consists of spotting a first array with large droplets of enzymes with varying concentrations and subsequently spotting a second array with small droplets of fluorogenic substrate on top of the enzyme array. By varying the array on array spotting patterns of different classes of enzyme (e.g., proteases, phosphatases, kinases) and their corresponding fluorogenic substrates, we have the unprecedented ability for testing enzymes and mixed samples in a multiplexed fashion within a single microarray slide. This new approach enables rapid enzyme characterization building upon a one enzyme on one slide droplet-based screening concept previously established.
authors.affiliation: Defence Medical and Environmental Research Institute, DSO National Laboratories, #09-01, 27 Medical Drive, Singapore, 117510, Singapore.
authors.forename: Brendan Fu-Long
authors.initials: BF
authors.lastname: Sieow
authors.affiliation: Defence Medical and Environmental Research Institute, DSO National Laboratories, #09-01, 27 Medical Drive, Singapore, 117510, Singapore. mahesh@dso.org.sg.|Department of Chemistry, National University of Singapore, Singapore, Singapore. mahesh@dso.org.sg.|Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore, Singapore. mahesh@dso.org.sg.
authors.forename: Mahesh
authors.initials: M
authors.lastname: Uttamchandani
mesh_terms: D001896:Boron Compounds / Q000737:chemistry; D002364:Caseins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000378:metabolism; D046228:Microarray Analysis; D010447:Peptide Hydrolases / Q000378:metabolism*; D040081:Protein Array Analysis / Q000379:methods*
chemical_list: C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D002364:Caseins; D005456:Fluorescent Dyes; D010447:Peptide Hydrolases
keywords: Activity-based screening; Array-on-array; Enzyme profiling; High-throughput screening; Small molecule arrays