A robust small-molecule microarray platform for screening cell lysates.

James E Bradner; Olivia M McPherson; Ralph Mazitschek; David Barnes-Seeman; John P Shen; Jasmeet Dhaliwal; Kristen E Stevenson; Jay L Duffner; Seung Bum Park; Donna S Neuberg; Paul Nghiem; Stuart L Schreiber; Angela N Koehler

doi:10.1016/j.chembiol.2006.03.004

PubMed · 16720270

A robust small-molecule microarray platform for screening cell lysates.

James E Bradner·Olivia M McPherson·Ralph Mazitschek·David Barnes-Seeman·John P Shen·Jasmeet Dhaliwal·Kristen E Stevenson·Jay L Duffner·Seung Bum Park·Donna S Neuberg·Paul Nghiem·Stuart L Schreiber·Angela N Koehler

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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.

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Cell Line Fluorescence Humans Oligonucleotide Array Sequence Analysis Tacrolimus Binding Protein 1A Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

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James E Bradner, Olivia M McPherson, Ralph Mazitschek, David Barnes-Seeman, John P Shen, Jasmeet Dhaliwal, Kristen E Stevenson, Jay L Duffner, Seung Bum Park, Donna S Neuberg, Paul Nghiem, Stuart L Schreiber, Angela N Koehler. 2006. A robust small-molecule microarray platform for screening cell lysates.. https://doi.org/10.1016/j.chembiol.2006.03.004

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