Label-free optical detection of small-molecule compound microarrays immobilized on solid support using macromolecular scaffolds and subsequent protein binding reactions

Y. S. Sun; J. P. Landry; Y. Y. Fei; X. D. Zhu; J. T. Luo; X. B. Wang; K. S. Lam

doi:10.1117/12.809636

12 February 2009 Label-free optical detection of small-molecule compound microarrays immobilized on solid support using macromolecular scaffolds and subsequent protein binding reactions

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

Author Affiliations +

Proceedings Volume 7182, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII; 71820P (2009) https://doi.org/10.1117/12.809636
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

Small-molecule microarrays composed of tens of thousands of distinct synthetic molecules, natural products, and their combinations/modifications provide a high-throughput platform for studying protein-ligand interactions. Immobilization of small molecule compounds on solid supports remains a challenge as widely varied small molecules generally lack unique chemical groups that readily react with singly or even multiply functionalized solid support. We explored two strategies for immobilizing small molecule compounds on epoxy-functionalized glass surface using primary-aminecontaining macromolecular scaffolds: bovine serum albumin (BSA) and amine-modified poly-vinyl alcohol (PVA). Small molecules with N-hydroxysuccinimide (NHS) groups were conjugated to BSA or amine-modified PVA. Small-molecule-BSA conjugates and small-molecule-PVA conjugates were subsequently immobilized on epoxy-functionalized glass slides through amine-epoxy reactions. Using an oblique-incidence reflectivity difference (OI-RD) scanning microscope as a label-free detector, we performed a comparative study of the effectiveness of BSA and PVA as macromolecular scaffolds for anchoring small molecule compounds in terms of conjugation efficiency, surface immobilization efficiency, effect of the scaffold on end-point and kinetics of subsequent binding reactions with protein probes.

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Y. S. Sun, J. P. Landry, Y. Y. Fei, X. D. Zhu, J. T. Luo, X. B. Wang, and K. S. Lam "Label-free optical detection of small-molecule compound microarrays immobilized on solid support using macromolecular scaffolds and subsequent protein binding reactions", Proc. SPIE 7182, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII, 71820P (12 February 2009); https://doi.org/10.1117/12.809636

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