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Sensors 2017, 17(1), 145; doi:10.3390/s17010145

Detection of the Vascular Endothelial Growth Factor with a Novel Bioluminescence Resonance Energy Transfer Pair Using a Two-Component System

Department of Ophthalmology, Justus-Liebig-University Giessen, Friedrichstr. 18, 35390 Giessen, Germany
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Author to whom correspondence should be addressed.
Academic Editors: Jong Seung Kim and Min Hee Lee
Received: 22 November 2016 / Revised: 30 December 2016 / Accepted: 9 January 2017 / Published: 13 January 2017
(This article belongs to the Special Issue Colorimetric and Fluorescent Sensor)
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Abstract

In this paper we describe a two-component BRET (bioluminescence resonance energy transfer)-based method to detect vascular endothelial growth factor (VEGF) molecules in unknown samples as the basis for subsequent in vivo use. A luminescent VEGF binding molecule, which binds in the receptor binding motif of VEGF, is used as the energy donor, transferred to a fluorophore-coupled VEGF binding molecule (acceptor), which binds to the neuropilin binding motif of VEGF, thus enabling energy transfer from the donor to the acceptor molecule. This leads to the emission of light at a longer wavelength and thus the generation of an increased BRET signal only when VEGF is bound to both the donor and acceptor molecules. We further describe a novel BRET pair that uses the Renilla reniformis mutant luciferase RLuc8 and the chemically engineered fluorophore PerCP-Cy5.5®, which exhibits superior peak separation of approximately 300 nm. The implantation of capsules consisting of the two BRET components in solution, permeable for VEGF for its in vivo detection, would provide a new and improved method for monitoring VEGF-induced pathologies and thus an adjustment of therapy to patient needs. View Full-Text
Keywords: VEGF; neuropilin; BRET; ranibizumab; single chain variable fragment (scFv); RLuc8; PerCP-Cy5.5® VEGF; neuropilin; BRET; ranibizumab; single chain variable fragment (scFv); RLuc8; PerCP-Cy5.5®
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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

Wimmer, T.; Schroeter, E.; Lorenz, B.; Stieger, K. Detection of the Vascular Endothelial Growth Factor with a Novel Bioluminescence Resonance Energy Transfer Pair Using a Two-Component System. Sensors 2017, 17, 145.

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