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 Citations (Scopus)

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

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Labels
Assays
incidence
proteins
Proteins
reflectance
Incidence
Surface Plasmon Resonance
Surface plasmon resonance
Biomolecules
surface plasmon resonance
Kinetics
interactions
kinetics
time signals
sensitivity
Photodiodes
Proteomics
photodiodes
Monolayers

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Real-time, label-free detection of biomolecular interactions in sandwich assays by the oblique-incidence reflectivity difference technique. / Sun, Yung Shin; Zhu, Xiangdong.

In: Sensors (Switzerland), Vol. 14, No. 12, 2014, p. 23307-23320.

Research output: Contribution to journalArticle

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