Fluospec platereader
Fluospec platereader is part of a drug discovery platform utilising fluorescent dye tags with long fluorescence lifetimes. Assays with fluorescence lifetime discrimination have a number of important advantages:
fast, convenient assay development with functionalised dye reagents
a range of enzyme substrates and ready-to-run assays for kinase and protease profiling
suppression of interference from coloured and fluorescent compounds
- improved rates of false positives and false negatives compared to simple intensity assays
VALIDATE reliably with the FLUOSPEC ® FL plate reader
Validating ‘hits’ from a primary high throughout compound screen is an important step in removing artefacts from datasets. Screening 1 million compounds at a hit rate of ~1% produces 10,000 validation tests before attractive looking leads can be selected. With such a large number of tests, each comprising perhaps 14 individual measurements, reliability and efficiency is crucial. A leading pharmaceutical company tested the reliability of IC50 values obtained with the FLUOSPEC ® FL under normal operating procedures in an automation lab. Excellent reliability was demonstrated.
Twelve point IC50 measurements were made for 394 compounds using a PURETIME ® SRC kinase assay (3µL final volume in 1536 well plate). Control wells were randomly distributed across the plate and each compound concentration was represented by a single well. Reliability was determined by comparing the derived IC50 values from a duplicate 1536 well plate. A correlation plot was used to compare the IC 50 values obtained from single well dose response series from the two plates. The FLUOSPEC ® FL produced highly correlated IC50 values over the whole concentration range from 0.1nM to 100µM, demonstrating reliable hit validation in an automation lab.
Tradeshow demonstration of Fluospec integrated with plate robot