Label-free screening small-molecule compound libraries for protein-ligands using a high-throughput optical scanning microscope

Y. Y. Fei, J. P. Landry, Y. S. Sun, X. D. Zhu, X. B. Wang, J. T. Luo, C. Y. Wu, Kit Lam

Research output: Chapter in Book/Report/Conference proceedingConference contribution

2 Scopus citations

Abstract

We describe a new oblique-incidence reflectivity difference (OI-RD) scanning microscope for high-throughput screening, in microarray format on functionalized glass slides, small-molecule compound libraries for protein ligands. The microscope employs a combination of scan mirror for y-scan and single-axis translation stage for x-scan. For a printed microarray with over 10,000 features, each of 100 ?m in diameter and distinct small molecule targets, we can acquire an end-point image of the microarray in 90 minutes with pixel resolution of 20 μm × 20μm. The microscope is also capable of measuring binding kinetics of over 10,000 protein-ligand reactions simultaneously. We also describe a number of strategies for immobilizing small molecule compounds on functionalized glass slides: (1) conjugating the compounds (through a chemically inert linker) with a lysine residue so that the primary amine on the lysine serves as the anchor to epoxy-functionalized glass surface; (2) conjugating the compounds (through a linker) with a biotin residue so that the biotin serves as the anchor to streptavidin-functionalized glass surface; (3) immobilizing small molecule compounds without modification on isocyanate-functionalized glass surface through non-specific reaction of nucleophilic molecular motifs on most bioactive compounds with isocyanate groups. We present preliminary measurements of protein-small molecule binding reactions using the new microscope and the surface immobilization strategies.

Original languageEnglish (US)
Title of host publicationProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume7182
DOIs
StatePublished - 2009
EventImaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII - San Jose, CA, United States
Duration: Jan 26 2009Jan 28 2009

Other

OtherImaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII
CountryUnited States
CitySan Jose, CA
Period1/26/091/28/09

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Keywords

  • High-throughput
  • Immobilization strategies
  • In situ detection
  • Label-free detection
  • Microarrays
  • Oblique-incidence reflectivity difference
  • OI-RD

ASJC Scopus subject areas

  • Applied Mathematics
  • Computer Science Applications
  • Electrical and Electronic Engineering
  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics

Cite this

Fei, Y. Y., Landry, J. P., Sun, Y. S., Zhu, X. D., Wang, X. B., Luo, J. T., Wu, C. Y., & Lam, K. (2009). Label-free screening small-molecule compound libraries for protein-ligands using a high-throughput optical scanning microscope. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE (Vol. 7182). [71820Q] https://doi.org/10.1117/12.809700