28 July 2000 Investigation of labeling FRET pairs to biomolecules for the development of dual-receptor biosensors
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We report herein on the development of a dual receptor detection method for enhanced biosensor monitoring. The proposed scheme requires the integration of a chemical transducer system with two unique protein receptors that bind to a single biological agent. Optical transduction occurs because the two protein receptors are tagged with special molecular groups. When bound to a single biological agent, these fluorescently labeled proteins undergo a change in fluorescence. This `fluorescent switching' relies on the well-known mechanisms of fluorescent resonance energy transfer (FRET). The paper focuses on the investigation and optimization of the chemical transduction system (FRET). A number of FRET dye pairs were tested in a spectrofluorimeter, and promising FRET pairs (FITC/TRITC and DMACA/FITC) were further tagged to the protein, avidin and its ligand, biotin. Due to their affinities, the FRET-tagged biomolecules combine in solution, resulting in a stable, fluorescent signal from the acceptor FRET dye with a simultaneous decrease in fluorescent signal from the donor FRET dye. The results indicate that the determined FRET pairs can be utilized in the development of dual receptor sensors.
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Sheila A. Grant, Sheila A. Grant, Juntao Xu, Juntao Xu, Edward J. Bergeron, Edward J. Bergeron, Jennifer Mroz, Jennifer Mroz, Amanda Rauschel, Amanda Rauschel, "Investigation of labeling FRET pairs to biomolecules for the development of dual-receptor biosensors", Proc. SPIE 4036, Chemical and Biological Sensing, (28 July 2000); doi: 10.1117/12.394059; https://doi.org/10.1117/12.394059


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