Comparison of static and microfluidic protease assays using modified bioluminescence resonance energy transfer chemistry.

ABSTRACT:

Background

Fluorescence and bioluminescence resonance energy transfer (F/BRET) are two forms of Förster resonance energy transfer, which can be used for optical transduction of biosensors. BRET has several advantages over fluorescence-based technologies because it does not require an external light source. There would be benefits in combining BRET transduction with microfluidics but the low luminance of BRET has made this challenging until now.

Methodology

We used a thrombin bioprobe based on a form of BRET (BRET(H)), which uses the BRET(1) substrate, native coelenterazine, with the typical BRET(2) donor and acceptor proteins linked by a thrombin target peptide. The microfluidic assay was carried out in a Y-shaped microfluidic network. The dependence of the BRET(H) ratio on

SUBMITTER: Wu N 

PROVIDER: S-EPMC3925127 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature