Measurement of drug-target engagement in live cells by two-photon fluorescence anisotropy imaging.

ABSTRACT: The ability to directly image and quantify drug-target engagement and drug distribution with subcellular resolution in live cells and whole organisms is a prerequisite to establishing accurate models of the kinetics and dynamics of drug action. Such methods would thus have far-reaching applications in drug development and molecular pharmacology. We recently presented one such technique based on fluorescence anisotropy, a spectroscopic method based on polarization light analysis and capable of measuring the binding interaction between molecules. Our technique allows the direct characterization of target engagement of fluorescently labeled drugs, using fluorophores with a fluorescence lifetime larger than the rotational correlation of the bound complex. Here we describe an optimized protocol

SUBMITTER: Vinegoni C 

PROVIDER: S-EPMC5928516 | biostudies-literature | 2017 Jul

REPOSITORIES: biostudies-literature