Research Papers: General

Re-evaluation of biotin-streptavidin conjugation in Förster resonance energy transfer applications

[+] Author Affiliations
Bahar Saremi

University of Texas at Arlington, Department of Bioengineering, Ultrasound and Optical Imaging Laboratory, Arlington, 500 UTA Boulevard, Texas 76010, United States

University of Texas at Arlington and University of Texas Southwestern Medical Center at Dallas, Joint Biomedical Engineering Program, Texas 75390, United States

Ming-Yuan Wei

University of Texas at Arlington, Department of Bioengineering, Ultrasound and Optical Imaging Laboratory, Arlington, 500 UTA Boulevard, Texas 76010, United States

University of Texas at Arlington and University of Texas Southwestern Medical Center at Dallas, Joint Biomedical Engineering Program, Texas 75390, United States

Yuan Liu

University of Texas at Arlington, Department of Bioengineering, Ultrasound and Optical Imaging Laboratory, Arlington, 500 UTA Boulevard, Texas 76010, United States

University of Texas at Arlington and University of Texas Southwestern Medical Center at Dallas, Joint Biomedical Engineering Program, Texas 75390, United States

Bingbing Cheng

University of Texas at Arlington, Department of Bioengineering, Ultrasound and Optical Imaging Laboratory, Arlington, 500 UTA Boulevard, Texas 76010, United States

University of Texas at Arlington and University of Texas Southwestern Medical Center at Dallas, Joint Biomedical Engineering Program, Texas 75390, United States

Baohong Yuan

University of Texas at Arlington, Department of Bioengineering, Ultrasound and Optical Imaging Laboratory, Arlington, 500 UTA Boulevard, Texas 76010, United States

University of Texas at Arlington and University of Texas Southwestern Medical Center at Dallas, Joint Biomedical Engineering Program, Texas 75390, United States

J. Biomed. Opt. 19(8), 085008 (Aug 27, 2014). doi:10.1117/1.JBO.19.8.085008
History: Received March 30, 2014; Revised July 29, 2014; Accepted August 6, 2014
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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.

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© 2014 Society of Photo-Optical Instrumentation Engineers

Citation

Bahar Saremi ; Ming-Yuan Wei ; Yuan Liu ; Bingbing Cheng and Baohong Yuan
"Re-evaluation of biotin-streptavidin conjugation in Förster resonance energy transfer applications", J. Biomed. Opt. 19(8), 085008 (Aug 27, 2014). ; http://dx.doi.org/10.1117/1.JBO.19.8.085008


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