title: A method for small molecule microarray-based screening for the rapid discovery of affinity-based probes.
abstract: We describe herein a new method for the high-throughput identification of affinity-based probes (AfBPs) using a small molecule microarray (SMM) approach. A hydroxylethylene-based small molecule library was first generated by solid-phase combinatorial synthesis. The library was tagged with biotin to facilitate immobilization on avidin-coated slides. Preliminary screening with γ-secretase (both the recombinantly purified protein as well as cellular lysates overexpressing the enzyme) was carried out, in order to identify potential small molecule binders, which were subsequently redesigned into AfBPs. Several specific and potent probes for γ-secretase were thus identified through the binding profiles observed on the SMMs. The SMM platform was able to sensitively and conveniently report activity-based binding interactions between aspartic proteases and their small molecule inhibitors. This new approach thus provides a potentially more rapid and efficient method for developing AfBPs using SMMs.
authors.affiliation: The NUS MedChem Program of the Office of Life Sciences, Faculty of Science, National University of Singapore, Singapore, Singapore.
authors.forename: Haibin
authors.initials: H
authors.lastname: Shi
authors.forename: Mahesh
authors.initials: M
authors.lastname: Uttamchandani
authors.forename: Shao Q
authors.initials: SQ
authors.lastname: Yao
mesh_terms: D053829:Amyloid Precursor Protein Secretases / Q000378:metabolism; D053000:Analytic Sample Preparation Methods; D000818:Animals; D057055:Aspartic Acid Proteases / Q000378:metabolism; D001360:Avidin / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005898:Glass / Q000737:chemistry; D057166:High-Throughput Screening Assays; D046228:Microarray Analysis / Q000379:methods*; D015335:Molecular Probes / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D054852:Small Molecule Libraries / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*
chemical_list: D015335:Molecular Probes; D054852:Small Molecule Libraries; D001360:Avidin; D053829:Amyloid Precursor Protein Secretases; D057055:Aspartic Acid Proteases