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Journal of Biomolecular Screening
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Article

Improving Lanthanide-Based Resonance Energy Transfer Detection by Increasing Donor-Acceptor Distances

Kurt W. Vogel* Kevin L. Vedvik

Invitrogen Drug Discovery Solutions, Madison, WI.

* To whom correspondence should be addressed. E-mail: kurt.vogel{at}invitrogen.com.


  Abstract

Lanthanide-based resonance energy transfer (LRET) is an established method for measuring or detecting proximity between a luminescent lanthanide (energy donor) and an organic fluorophore (energy acceptor). Because resonance energy transfer is a distance-dependent phenomenon that increases in efficiency to the 6th power of the distance between the donor and the acceptor, assay systems are often designed to minimize donor-acceptor distances. However, the authors show that because of the R6 relationship between transfer efficiency and sensitized emission lifetime, energy transfer can be difficult to measure in a time-gated manner when the donor-acceptor distance is small relative to the Förster radius. In such systems, the advantages inherent in time-resolved, ratiometric measurements are lost but can be regained by designing the system such that the average donor-acceptor distance is increased.

Key Words: TR-FRET, lanthanides, protease, assay

First published on April 28, 2006, doi:10.1177/1087057106287142

Journal of Biomolecular Screening 2006;11:439.

A more recent version of this article appeared on June 1, 2006

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