A robust small-molecule microarray platform for screening cell lysates.
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.
Bradner
James E
JE
Broad Institute of Harvard and MIT, 7 Cambridge Center, Cambridge, Massachusetts 02142, USA.
McPherson
Olivia M
OM
Mazitschek
Ralph
R
Barnes-Seeman
David
D
Shen
John P
JP
Dhaliwal
Jasmeet
J
Stevenson
Kristen E
KE
Duffner
Jay L
JL
Park
Seung Bum
SB
Neuberg
Donna S
DS
Nghiem
Paul
P
Schreiber
Stuart L
SL
Koehler
Angela N
AN
D002460:Cell Line; D005453:Fluorescence; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D022061:Tacrolimus Binding Protein 1A / Q000737:chemistry
D022061:Tacrolimus Binding Protein 1A