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, Kit Lam

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

2 Scopus citations

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. Smallmolecule-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.

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

Keywords

  • Ellipsometry
  • Equilibrium dissociation constant
  • Kinetics
  • Label-free detection
  • Langmuir two-site model
  • Oblique-incidence reflectivity difference
  • OIRD
  • Protein-ligand interaction
  • Small-molecule microarrays

ASJC Scopus subject areas

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

Fingerprint Dive into the research topics of 'Label-free optical detection of small-molecule compound microarrays immobilized on solid support using macromolecular scaffolds and subsequent protein binding reactions'. Together they form a unique fingerprint.

Cite this