Real-time, label-free detection of biomolecular interactions in sandwich assays by the oblique-incidence reflectivity difference technique

Yung Shin Sun, Xiangdong Zhu

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

One of the most important goals in proteomics is to detect the real-time kinetics of diverse biomolecular interactions. Fluorescence, which requires extrinsic tags, is a commonly and widely used method because of its high convenience and sensitivity. However, in order to maintain the conformational and functional integrality of biomolecules, label-free detection methods are highly under demand. We have developed the oblique-incidence reflectivity difference (OI-RD) technique for label-free, kinetic measurements of protein-biomolecule interactions. Incorporating the total internal refection geometry into the OI-RD technique, we are able to detect as low as 0.1% of a protein monolayer, and this sensitivity is comparable with other label-free techniques such as surface plasmon resonance (SPR). The unique advantage of OI-RD over SPR is no need for dielectric layers. Moreover, using a photodiode array as the detector enables multi-channel detection and also eliminates the over-time signal drift. In this paper, we demonstrate the applicability and feasibility of the OI-RD technique by measuring the kinetics of protein-protein and protein-small molecule interactions in sandwich assays.

Original languageEnglish (US)
Pages (from-to)23307-23320
Number of pages14
JournalSensors (Switzerland)
Volume14
Issue number12
DOIs
StatePublished - 2014

ASJC Scopus subject areas

  • Medicine(all)

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