Ionic Current measurements

Whole-cell current amplitude and kinetics measurements verify the results of the interactions of a test article with a selected ionic current. The assay can be used to elucidate the mechanism behind the block of ion channels by a test article, for instance, or to confirm the nature of a target when ion-channel inhibition is a suspected part of the efficacious mechanism. The experiments are conducted on heterologous transfect ion systems, freshly isolated (primary cultures) cells, or specifically prepared tissues, e.g.: brain slices

Method

Manual patch-clamp, whole-cell configuration

HEK293, LNCaP, CHO or isolated cardiac cells

• Number of cells exposed to the test article: 7 (screens: 3)
• Number of concentrations of test article: 4 (screens: 3)
• Method of test article exposure: cumulative concentrations in closed-circuit perfusion, or open-ended perfusion of individual concentrations
• Voltage pulse protocols customized based on the selected ionic current to measure
• A study includes positive, negative, and vehicle controls

• A quantitative assessment of ionic current inhibition, type of block, etc.
• Determination of the IC50 value (if applicable)
• Analysis of both amplitude and current kinetics as indicators of inhibition
• FDA-ready hard copy  and e-report for electronic IND submission
• Holistic interpretation of a positive signal, considering all other data generated

Pre-IND (GLP-compliant) or exploratory designs (screens are non-GLP compliant)