Direct detection of aptamer-thrombin binding via surface-enhanced Raman spectroscopy

Cynthia V. Pagba, Stephen M. Lane, Hansang Cho, Sebastian Wachsmann-Hogiu

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

In this study, we exploit the sensitivity offered by surfaceenhanced Raman scattering (SERS) for the direct detection of thrombin using the thrombin-binding aptamer (TBA) as molecular receptor. The technique utilizes immobilized silver nanoparticles that are functionalized with thiolated thrombin-specific binding aptamer, a 15-mer (5'-GGTTGGTGTGGTTGG-3') quadruplex forming oligonucleotide. In addition to the Raman vibrational bands corresponding to the aptamer and blocking agent, new peaks (mainly at 1140, 1540, and 1635 cm-1) that are characteristic of the protein are observed upon binding of thrombin. These spectral changes are not observed when the aptamer-nanoparticle assembly is exposed to a nonbinding protein such as bovine serum albumin (BSA). This methodology could be further used for the development of label-free biosensors for direct detection of proteins and other molecules of interest for which aptamers are available.

Original languageEnglish (US)
Article number047006
JournalJournal of Biomedical Optics
Volume15
Issue number4
DOIs
StatePublished - Jul 2010

Keywords

  • Aptamer
  • Direct detection
  • Label-free assay
  • Quadruplex
  • Surface-enhanced raman scattering
  • Thrombin

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biomaterials
  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics

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