SYLICA Lecture: Fluorescence polarization immunoassays for simple, high-throughput screening of small molecules

WHEN: 16/12/2014,11:00
WHERE: room 205, building A11, Masaryk University Campus in Bohunice, Kamenice 5, Brno

Abstract:
The control for level of environmental and food contamination must be in global scale with high-throughput screening (HTS). The current methods of assays like chromatography techniques have several limitations for HTS. The immunochemical methods based on the specific and high affinity antibodies are widely used for high-throughput screening and monitoring of many physiologies active compounds. The method of fluorescence polarization immunoassay (FPIA) could be used for rapid determination of small molecules. FPIA is based on the competition of an analyte and the analyte labeled with a fluorescent label (tracer) for the binding sites of specific antibodies. In the absence of analytes in the sample the determined fluorescence polarization is a maximum and decreases with increasing concentration of analyte. The total time required for an assay is few seconds or minutes. The limit of detection for analyte could be around 10 ng/ml after careful optimization antibodies and fluorescein-labeled tracers. FPIA is simple method in the formulation and allows for a few minutes with both qualitative and quantitative determination of small molecules. The FPIA is an accurate and cost-efficient method which is readily automated and suitable for quantitative detection of multi analytes in one sample. The recent results for development of FPIA for dyes and drugs will be presented. More details about this FPIA method can be found in the reviews and in some of our recent publications.