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; K. S. Lam

doi:10.1117/12.809700

12 February 2009 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, K. S. Lam

Author Affiliations +

Proceedings Volume 7182, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII; 71820Q (2009) https://doi.org/10.1117/12.809700
Event: SPIE BiOS, 2009, San Jose, California, United States

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 μm 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 × 20 μm. 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.

Citation Download Citation

Y. Y. Fei, J. P. Landry, Y. S. Sun, X. D. Zhu, X. B. Wang, J. T. Luo, C. Y. Wu, and K. S. Lam "Label-free screening small-molecule compound libraries for protein-ligands using a high-throughput optical scanning microscope", Proc. SPIE 7182, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII, 71820Q (12 February 2009); https://doi.org/10.1117/12.809700

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