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Sensors 2014, 14(12), 23307-23320; doi:10.3390/s141223307

Real-Time, Label-Free Detection of Biomolecular Interactions in Sandwich Assays by the Oblique-Incidence Reflectivity Difference Technique

1
Department of Physics, Fu-Jen Catholic University, New Taipei City 24205, Taiwan
2
Department of Physics, University of California at Davis, Davis, CA 95616, USA
*
Author to whom correspondence should be addressed.
Received: 21 October 2014 / Revised: 20 November 2014 / Accepted: 25 November 2014 / Published: 5 December 2014
(This article belongs to the Special Issue Advances in Optical Biosensors)
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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. View Full-Text
Keywords: label-free biosensor; oblique-incidence reflectivity difference (OI-RD); protein-protein interaction; protein-small molecule interaction; real-time kinetics label-free biosensor; oblique-incidence reflectivity difference (OI-RD); protein-protein interaction; protein-small molecule interaction; real-time kinetics
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Sun, Y.-S.; Zhu, X. Real-Time, Label-Free Detection of Biomolecular Interactions in Sandwich Assays by the Oblique-Incidence Reflectivity Difference Technique. Sensors 2014, 14, 23307-23320.

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