Optimizing the hERG Assay on the PatchXpress 7000A System for Compounds that Demonstrate Non-specific Binding
Poster Dec 22, 2005
Iris Yang, Naibo Yang, Cathy Smith-Maxwell, Claire Quinn, Kirsty Macfarlane and David Yamane
AbstractNon-specific binding of compound can affect the potency measurement from both conventional and automated patch-clamp assays. Because of this, pre-cautions need to be taken to ensure this is minimized or eliminated. Eight compounds, astemizole, pimozide, dofetilide, cisapride, terfenadine, flunarizine, quinidine and imipramine were used to evaluate the effect of different assay conditions to overcome the effects of non-specific binding in our hERG assay on the PatchXpress 7000A system.
To achieve the best IC50 values, in addition to optimizing instrument parameters to minimize non-specific binding, care had to be taken in all phases of compound preparation. Compound binding to plastic container need to be minimized. Buffers used for the assay also need to be optimized. In this poster, we present the assay conditions found that generated the best IC50 values on the PatchXpress 7000A as well as the assay results for these compounds.
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE
During early drug discovery, the study of metabolism plays an essential role in determining which drug candidates move forward into development and later stages. As an alternative to traditional Data Dependent Acquisition (DDA), the use of MSE/All Ions Fragmentation (AIF) has become common in metabolite identification workflows for the analysis of metabolic hot spots. Here we present a solution for analysis of MSE/AlF in metID studies.READ MORE