OpAns metabolic stability assays (liver microsomes and S9) assist in metabolic profiling for lead optimization. We provide non-cellular assays that utilize highly specific LC-MS or LC-MS/MS detection. Results can be returned in standardized or custom reports to facilitate integration with your data management tools.
Standard Assay:
Stock solutions are prepared from the dry powder. Incubation wells are assembled from protein preparations, cofactors and diluted compound stocks. The time zero sample is taken immediately after assembly. Subsequent samples are taken at specified timepoints (e.g. 15, 30, 60 min). Quantification is performed on a LC-MS/MS system (Sciex API3000). A ratio is made between the response of the incubated samples and time zero to determine the percent parent compound remaining. All assays are performed in triplicate and the average percent parent compound remaining is reported. The whole assay (assembly, incubation, sampling) is automated on a Tecan robotic system.
High-Throughput Assay:
The customer provides DMSO stocks in either 96 or 384 well microtiter plates. The DMSO stocks are serially diluted with buffer to a concentration suitable for the assay. The exact assay assembly and sampling protocol is usually optimized depending on the specific requirements (96 or 384 well, number of species, number of timepoints). Quantification is performed on an Agilent 1100 LC-MSD system with the multi-mode source. A ratio is made between the response of the incubated samples and time zero to determine the percent parent compound remaining. All assays are performed in duplicate and the average percent parent compound remaining is reported.
