title: A robust small-molecule microarray platform for screening cell lysates.
abstract: Herein we report the expanded functional group compatibility of small-molecule microarrays to include immobilization of primary alcohols, secondary alcohols, phenols, carboxylic acids, hydroxamic acids, thiols, and amines on a single slide surface. Small-molecule "diversity microarrays" containing nearly 10,000 known bioactive small molecules, natural products, and small molecules originating from several diversity-oriented syntheses were produced by using an isocyanate-mediated covalent capture strategy. Selected printed bioactive compounds were detected with antibodies against compounds of interest. The new surface of the diversity microarrays is highly compatible with approaches involving cellular lysates. This feature has enabled a robust, optimized screening methodology using cellular lysates, allowing the detection of specific interactions with a broad range of binding affinity by using epitope-tagged or chimeric fluorescent proteins without prior purification. We believe that this expanded research capability has considerable promise in biology and medicine.
authors.affiliation: Broad Institute of Harvard and MIT, 7 Cambridge Center, Cambridge, Massachusetts 02142, USA.
authors.forename: James E
authors.initials: JE
authors.lastname: Bradner
authors.forename: Olivia M
authors.initials: OM
authors.lastname: McPherson
authors.forename: Ralph
authors.initials: R
authors.lastname: Mazitschek
authors.forename: David
authors.initials: D
authors.lastname: Barnes-Seeman
authors.forename: John P
authors.initials: JP
authors.lastname: Shen
authors.forename: Jasmeet
authors.initials: J
authors.lastname: Dhaliwal
authors.forename: Kristen E
authors.initials: KE
authors.lastname: Stevenson
authors.forename: Jay L
authors.initials: JL
authors.lastname: Duffner
authors.forename: Seung Bum
authors.initials: SB
authors.lastname: Park
authors.forename: Donna S
authors.initials: DS
authors.lastname: Neuberg
authors.forename: Paul
authors.initials: P
authors.lastname: Nghiem
authors.forename: Stuart L
authors.initials: SL
authors.lastname: Schreiber
authors.forename: Angela N
authors.initials: AN
authors.lastname: Koehler
mesh_terms: D002460:Cell Line; D005453:Fluorescence; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D022061:Tacrolimus Binding Protein 1A / Q000737:chemistry
chemical_list: D022061:Tacrolimus Binding Protein 1A