DEVELOPMENT OF A SENSITIVE INSTRUMENT FOR BIOLUMINESCENCE RESONANCE ENERGY TRANSFER (BRET) APPLICATIONS

摘要

The instrument developed has successfully proven to be a sensitive and very suitable solution for assays based on BRET technology. Table 1 clearly shows the improvement by using a dedicated luminescence measurement path compared to a fluorescence measurement path. Table 1. Comparison of relative sensitivity and cross-talk between the luminescence and fluorescence measurement position and a standard fluorometer based on a Dioxetane chemiluminescence standard (Std 1, Mediators Diagnostica, Vienna) Similar results have been derived from comparisons with other instruments which are basically modified fluorometers. The developed instrument is significantly superior to the existing modified fluorometers which have been used for that purpose so far. Besides BRET the instrument is applicable to a multitude of other luminescence and fluorescence assays, for example, the determination of receptor expression. The sensitivity of the instrument together with the stability of the signals will enable BRET to be used in compound screening as well as in basic research. In 7TM receptor screening a potentially universal functional assay for all types of receptors regardless of their stimulating or inhibiting nature has been established.