Detection of protein-small molecule binding using a self-referencing external cavity laser biosensor

Meng Zhang, Jessie Peh, Paul J. Hergenrother, Brian T. Cunningham

Research output: Contribution to journalArticle

Abstract

High-throughput screening has enabled the identification of small molecule modulators of important drug targets via well-established colorimetric or fluorimetric activity assays. However, existing methods to identify small molecule binders of nonenzymatic protein targets lack either the simplicity (e.g., require labeling one of the binding partners with a reporter) or throughput inherent in enzymatic assays widely used for HTS. Thus, there is intense interest in the development of high-throughput technologies for label-free detection of protein-small molecule interactions. Here we describe a novel self-referencing external cavity laser (ECL) biosensor approach that achieves high resolution and high sensitivity, while eliminating thermal noise with subpicometer wavelength accuracy. Using the self-referencing ECL biosensor, we demonstrate detection of binding between small molecules and a variety of immobilized protein targets, pairs that have binding affinities or inhibition constants ranging from subnanomolar to low micromolar. Finally, a "needle-in-the-haystack" screen for inhibitors against carbonic anhydrase isozyme II is performed, in which known inhibitors are clearly differentiated from inactive molecules within a compound library.

Original languageEnglish (US)
Pages (from-to)5840-5843
Number of pages4
JournalJournal of the American Chemical Society
Volume136
Issue number16
DOIs
StatePublished - Apr 23 2014

Fingerprint

Laser resonators
Biosensing Techniques
Biosensors
cavity
Lasers
laser
Immobilized Proteins
Carbonic Anhydrase II
Proteins
Molecules
protein
inhibitor
Enzyme Assays
assay
Isoenzymes
Libraries
Needles
Throughput
Hot Temperature
Assays

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Detection of protein-small molecule binding using a self-referencing external cavity laser biosensor. / Zhang, Meng; Peh, Jessie; Hergenrother, Paul J.; Cunningham, Brian T.

In: Journal of the American Chemical Society, Vol. 136, No. 16, 23.04.2014, p. 5840-5843.

Research output: Contribution to journalArticle

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