Bioanalytical systems based on bioluminescence resonance energy transfer using firefly luciferase

Bioanalytical systems based on the Bioluminescence Resonance Energy Transfer (BRET) are widely used in fundamental biochemical studies, as well as for screening and analysis of biologically active compounds. The Renilla luciferase is the most often used energy donor in this system despite the fact that it has low bioluminescence quantum yield and demonstrates not so stable luminescence in time as the firefly luciferase. Moreover, the bioluminescence λmax is observed in the green region of the spectrum, which complicates signal recording in tissues during in vivo experiments. The firefly luciferases do not have such drawbacks and show great promise for applications in BRET systems. Different versions of BRET systems based on firefly luciferases and the methods for increasing their efficiency are considered in this review; examples of the use of BRET systems based on the firefly luciferases for highly sensitive determination of proteases and for homogeneous immunoassays are presented.