FRET-based biosensors to detect infectious agents

Juntao Xu; Sheila A. Grant

doi:10.1117/12.456061

14 February 2002 FRET-based biosensors to detect infectious agents

Juntao Xu, Sheila A. Grant

Proceedings Volume 4578, Fiber Optic Sensor Technology and Applications 2001; (2002) https://doi.org/10.1117/12.456061
Event: Environmental and Industrial Sensing, 2001, Boston, MA, United States

Abstract

We report herein on the development of a FRET-based method to detect changes caused by viral protein-receptor binding. FRET fluorophore pairs (donor and acceptor fluorophores) were tagged to two specific receptors, both which bind to a viral protein. When the binding event occurs, the distance between the donor and acceptor FRET fluorophores is decreased, thus initiating the fluorescence resonance energy transfer (FRET). Since the binding event is unique to the viral protein, fluorescent change indicates the present of the virus. In this paper, the viral protein gp120, which is the featured protein on the surface of HIV-1, was detected. The receptors, CD4 and gp120-antibody which specifically bind to gp120, were conjugated to the FRET fluorophore pair, AMCA-NHS (succinimidyl-7-amino-4-methylcoumarin-3-acetic acid) and FITC (fluorescein isothiocyanate) respectively. Spectrofluorimetry was used to detect the fluorescent change between AMCA-NHS and FITC peak intensities when the receptors bind to the gp120. Specific binding gp120 and non-specific binding gp120 were used to test the selectivity of the sensor. The results indicated that FRET-conjugated receptors can efficiently detect the presence of gp120.

Citation Download Citation

Juntao Xu and Sheila A. Grant "FRET-based biosensors to detect infectious agents", Proc. SPIE 4578, Fiber Optic Sensor Technology and Applications 2001, (14 February 2002); https://doi.org/10.1117/12.456061

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