FRET based biosensor for detection of active NF-kB

Francesco Baldini; Lorenzo Citti; Claudio Domenici; Ambra Giannetti; Lorena Tedeschi; Tuan Vo-Dinh; Musundi B. Wabuyele

doi:10.1117/12.623696

23 May 2005 FRET based biosensor for detection of active NF-kB

Francesco Baldini, Lorenzo Citti, Claudio Domenici, Ambra Giannetti, Lorena Tedeschi, Tuan Vo-Dinh, Musundi B. Wabuyele

Proceedings Volume 5855, 17th International Conference on Optical Fibre Sensors; (2005) https://doi.org/10.1117/12.623696
Event: 17th International Conference on Optical Fibre Sensors, 2005, Bruges, Belgium

Abstract

The Nuclear Factor kB is a transcription factor, ubiquitously expressed, involved in the regulation of a large number of genes and in a variety of human disease including inflammation, asthma, atherosclerosis, AIDS, septic shock, arthritis and cancer. The critical need for a simple and direct method to evaluate the quantity of active NF-kB in a biological sample can be addressed using a suitable and reusable biosensor. For this purpose, a novel method, using fluorescence resonance energy transfer (FRET), to detect the active form of NF-kB binding a specific DNA sequence has been developed. A single-stranded DNA (ssDNA) with auto-complementary sequence has been properly designed and synthesized. In order to evaluate FRET due to the DNA/protein binding interaction taking place between double-stranded DNA (dsDNA) immobilized in a capillary wall and NF-kB proteins, a highly sensitive FRET-based biosensor system developed in our laboratory was used. Preliminary results show that our system was capable of detecting the active form of NF-kB protein with a detection efficiency of about 90% and that the system has a good regenerability.

Citation Download Citation

Francesco Baldini, Lorenzo Citti, Claudio Domenici, Ambra Giannetti, Lorena Tedeschi, Tuan Vo-Dinh, and Musundi B. Wabuyele "FRET based biosensor for detection of active NF-kB", Proc. SPIE 5855, 17th International Conference on Optical Fibre Sensors, (23 May 2005); https://doi.org/10.1117/12.623696

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