A Three-Stage Experimental Strategy to Evaluate and Validate an Interplate IC₅₀ Format

Abstract

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.

ICECAP: An Integrated, General-Purpose, Automation-Assisted IC50/EC50 Assay Platform

“…In the HTS field, there have been many reports of using new technologies, including automation, to increase IC 50 /EC 50 assay throughput. [12][13][14][15][16][17] From a MS-centric bioanalytical/biomarker lab's point of view, for each individual compound, there is nothing in the IC 50 /EC 50 experimental procedure that cannot be automated with current state-of-the-art bioanalytical automated sample preparation systems. 18,19 However the challenge lies in the overall production throughput.…”