FRET based biosensor for detection of active NF-kB


Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Baldini, F; Citti, L; Domenici, C; Giannetti, A; Tedeschi, L; Vo-Dinh, T; Wabuyele, MB

Published Date

  • December 13, 2005

Published In

Volume / Issue

  • 5855 PART I /

Start / End Page

  • 439 - 442

International Standard Serial Number (ISSN)

  • 0277-786X

Digital Object Identifier (DOI)

  • 10.1117/12.623696

Citation Source

  • Scopus