A three-stage experimental strategy to evaluate and validate an interplate IC50 format.
The serial dilution of compounds to establish potency against target enzymes or receptors can at times be a rate-limiting step in project progression. We have investigated the possibility of running 50% inhibitory concentration experiments in an interplate format, with dose ranges constructed across plates. The advantages associated with this format include a faster reformatting time for the compounds while also increasing the number of doses that can be potentially generated. These two factors, in particular, would lend themselves to a higher-throughput and more timely testing of compounds, while also maximizing chances to capture fully developed dose-response curves. The key objective from this work was to establish a strategy to assess the feasibility of an interplate format to ensure that the quality of data generated would be equivalent to historical formats used. A three-stage approach was adopted to assess and validate running an assay in an interplate format, compared to an intraplate format. Although the three-stage strategy was tested with two different assay formats, it would be necessary to investigate the feasibility for other assay types. The recommendation is that the three-stage experimental strategy defined here is used to assess feasibility of other assay formats used.
High Throughput Screening Centre of Emphasis, Worldwide Research, Sandwich, UK. dan.rodrigues@pfizer.com
Daniel J
DJ
Rodrigues
Richard
R
Lyons
Philip
P
Laflin
Wayne
W
Pointon
Juha
J
Kammonen
D003627:Data Interpretation, Statistical; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation / Q000379:methods*; D017930:Genes, Reporter / Q000235:genetics; D007201:Indicator Dilution Techniques; D015203:Reproducibility of Results; D012588:Scintillation Counting; D001618:beta-Lactamases / Q000235:genetics
D001618:beta-Lactamases