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High-throughput screening assay for the tunable selection of protein ligands.

Publication ,  Journal Article
Powell, KD; Fitzgerald, MC
Published in: Journal of combinatorial chemistry
March 2004

Here, we describe a new protein-ligand binding assay that is amenable to high-throughput screening applications. The assay involves the use of SUPREX (stability of unpurified proteins from rates of H/D exchange), a new H/D exchange and mass spectrometry-based technique we recently developed for the quantitative analysis of protein-ligand binding interactions. As part of this work, we describe a new high-throughput SUPREX protocol, and we demonstrate that this protocol can be used to efficiently screen peptide ligands in a model combinatorial library for binding to a model protein system, the S-protein. The high-throughput SUPREX protocol developed here is generally applicable to a wide variety of protein ligands, including DNA, small molecules, metals, and other proteins. On the basis of the results of the model study in this work, one person with access to one MALDI mass spectrometer should be able to screen approximately 10 000 compounds per 24-h period using the protocol described here. With full automation and the use of a commercially available MALDI mass spectrometer optimized for high-throughput analyses, we estimate that the SUPREX-based assay described here could be used to screen on the order of 100 000 ligands per day.

Duke Scholars

Published In

Journal of combinatorial chemistry

DOI

EISSN

1520-4774

ISSN

1520-4766

Publication Date

March 2004

Volume

6

Issue

2

Start / End Page

262 / 269

Related Subject Headings

  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Proteins
  • Protein Binding
  • Peptides
  • Organic Chemistry
  • Ligands
  • Hydrogen
  • Deuterium
  • Automation
  • 0305 Organic Chemistry
 

Citation

APA
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ICMJE
MLA
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Powell, K. D., & Fitzgerald, M. C. (2004). High-throughput screening assay for the tunable selection of protein ligands. Journal of Combinatorial Chemistry, 6(2), 262–269. https://doi.org/10.1021/cc034051e
Powell, Kendall D., and Michael C. Fitzgerald. “High-throughput screening assay for the tunable selection of protein ligands.Journal of Combinatorial Chemistry 6, no. 2 (March 2004): 262–69. https://doi.org/10.1021/cc034051e.
Powell KD, Fitzgerald MC. High-throughput screening assay for the tunable selection of protein ligands. Journal of combinatorial chemistry. 2004 Mar;6(2):262–9.
Powell, Kendall D., and Michael C. Fitzgerald. “High-throughput screening assay for the tunable selection of protein ligands.Journal of Combinatorial Chemistry, vol. 6, no. 2, Mar. 2004, pp. 262–69. Epmc, doi:10.1021/cc034051e.
Powell KD, Fitzgerald MC. High-throughput screening assay for the tunable selection of protein ligands. Journal of combinatorial chemistry. 2004 Mar;6(2):262–269.

Published In

Journal of combinatorial chemistry

DOI

EISSN

1520-4774

ISSN

1520-4766

Publication Date

March 2004

Volume

6

Issue

2

Start / End Page

262 / 269

Related Subject Headings

  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Proteins
  • Protein Binding
  • Peptides
  • Organic Chemistry
  • Ligands
  • Hydrogen
  • Deuterium
  • Automation
  • 0305 Organic Chemistry