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Fluorescent Signal Amplification of Carbocyanine Dyes Using Engineered Viral Nanoparticles

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Contribution from the Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, 4555 Overlook Avenue SW, Washington, D.C. 20375, Nova Research Inc., Alexandria, Virginia, 22308, and Department of Molecular Biology, Center for Interactive Molecular Biosciences, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California, 92037
Cite this: J. Am. Chem. Soc. 2006, 128, 15, 5184–5189
Publication Date (Web):March 25, 2006
https://doi.org/10.1021/ja058574x
Copyright © 2006 American Chemical Society

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    Abstract

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    We report enhancement in the fluorescent signal of the carbocyanine dye Cy5 by using an engineered virus as a scaffold to attach >40 Cy5 reporter molecules at fixed locations on the viral capsid. Although cyanine dye loading is often accompanied by fluorescence quenching, our results demonstrate that organized spatial distribution of Cy5 reporter molecules on the capsid obviates this commonly encountered problem. In addition, we observe energy transfer from the virus to adducted dye molecules, resulting in a highly fluorescent viral nanoparticle. We have used this enhanced fluorescence for the detection of DNA−DNA hybridization. When compared with the most often used detection methods in a microarray-based genotyping assay for Vibrio cholerae O139, these viral nanoparticles markedly increased assay sensitivity, thus demonstrating their applicability for existing DNA microarray protocols.

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    UV−visible spectrum of EF virus, additional fluorescence and excitation spectra, a table describing detailed preparation of NeutrAvidin−dye−virus series, and a table of gene targets, probes, and primer sequences. This material is available free of charge via the Internet at http://pubs.acs.org.

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