Fluorescence polarization immunoassays and related methods for simple, high-throughput screening of small molecules.
Fluorescence polarization immunoassay (FPIA) is a homogeneous (without separation) competitive immunoassay method based on the increase in fluorescence polarization (FP) of fluorescent-labeled small antigens when bound by specific antibody. The minimum detectable quantity of FPIAs with fluorescein label (about 0.1 ng analyte) is comparable with chromatography and ELISA methods, although this may be limited by sample matrix interference. Because of its simplicity and speed, FPIA is readily automated and therefore suitable for high-throughput screening (HTS) in a variety of application areas. Systems that involve binding of ligands to receptor proteins are also susceptible to analysis by analogous FP methods employing fluorescent-labeled ligand and HTS applications have been developed, notably for use in candidate drug screening.
Smith
David S
DS
MicroPharm Ltd, 51-53 Bartholomew Close, St. Bartholomew's Hospital, London, EC1A 7BE, UK.
Eremin
Sergei A
SA
D001322:Autoanalysis; D001667:Binding, Competitive; D004353:Drug Evaluation, Preclinical; D004784:Environmental Monitoring / Q000379:methods; D005452:Fluoresceins / Q000737:chemistry; D005454:Fluorescence Polarization / Q000379:methods*; D016231:Fluorescence Polarization Immunoassay / Q000379:methods*; D005455:Fluorescent Antibody Technique / Q000379:methods; D005456:Fluorescent Dyes / Q000737:chemistry; D005504:Food Analysis / Q000379:methods; D008024:Ligands; D036103:Nanotechnology / Q000379:methods*; D011506:Proteins / Q000032:analysis / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013194:Staining and Labeling
D005452:Fluoresceins; D005456:Fluorescent Dyes; D008024:Ligands; D011506:Proteins