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Journal of Biomedical Optics

Re-evaluation of biotin-streptavidin conjugation in Förster resonance energy transfer applications
Author(s): Bahar Saremi; Ming-Yuan Wei; Yuan Liu; Bingbing Cheng; Baohong Yuan
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Paper Abstract

Bioaffinity conjugation between streptavidin (SA) and biotin has been widely used to link donors and acceptors for investigating the distance-dependent Förster resonance energy transfer (FRET). When studying a commonly used FRET system of (QD-SA)-(biotin-DNA-dye) [donor: quantum dot (QD); acceptor: small organic fluorescent dye; and linker: deoxyribose nucleic acid (DNA) molecule via SA-biotin conjugation], however, a contradictory finding was recently reported in the literature. It was found that the FRET lost its dependence on the number of DNA base pairs when using a phosphate-buffered saline (PBS) solution. We found that the conflicted results were caused by the ionic strength of the adopted buffer solutions. Our results suggest that the dependent FRET on the number of DNA bases is favorable in a low-ionic-strength buffer, whereas in relatively high-ionic-strength buffers, the FRET loses the DNA length dependence. We propose that the independence is mainly caused by the conformational change of DNA molecules from a stretched to a coiled mode when the cations in the high-ionic-strength buffer neutralize the negatively charged backbone of DNA molecules, thereby bringing the acceptors close to the donors.

Paper Details

Date Published: 27 August 2014
PDF: 12 pages
J. Biomed. Opt. 19(8) 085008 doi: 10.1117/1.JBO.19.8.085008
Published in: Journal of Biomedical Optics Volume 19, Issue 8
Show Author Affiliations
Bahar Saremi, The Univ. of Texas at Arlington (United States)
The Univ. of Texas Southwestern Medical Ctr. at Dallas (United States)
Ming-Yuan Wei, The Univ. of Texas at Arlington (United States)
The Univ. of Texas Southwestern Medical Ctr. at Dallas (United States)
Yuan Liu, The Univ. of Texas at Arlington (United States)
The Univ. of Texas Southwestern Medical Ctr. at Dallas (United States)
Bingbing Cheng, The Univ. of Texas at Arlington (United States)
The Univ. of Texas Southwestern Medical Ctr. at Dallas (United States)
Baohong Yuan, The Univ. of Texas at Arlington (United States)
The Univ. of Texas Southwestern Medical Ctr. at Dallas (United States)


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