Patent ID: 8846575
Filing Date: 2014-09-30
Classification: C12Q,G01N

Abstract:
1. A method for hit compound identification in a high throughput, label-free biosensor cellular assay, the method comprising: (a) generating at least one signaling pathway-specific kinetic profile for a target cell membrane receptor in a live-cell, wherein the at least one signaling pathway-specific kinetic profile for the target cell membrane receptor is obtained by: (i) providing an optical biosensor having a single type of live-cell immobilized on a surface of the biosensor, the biosensor being situated in a well of a microtiter plate, and the live-cell having a first and second target, wherein the first target is a cell membrane receptor, wherein the second target is a cellular protein, wherein the cellular protein is a member of the signaling pathway triggered by activation of the cell membrane receptor, (ii) separately stimulating the live-cell expressing target cell membrane receptor with a plurality of candidate compounds after the live-cell is attached to the surface of the biosensor; (iii) contacting the ligand candidate-treated live-cell with a mixture containing two markers; and (iv) determining the effect of the ligand candidate on the marker mixture-induced biosensor output; (b) generating a relational table, which table incorporates each signaling pathway-specific kinetic profile for the plurality of candidate compounds; (c) determining a delta response based on the relational table for each of the plurality of candidate compounds screened against the target cell membrane receptor; and (d) identifying hit compounds from the plurality of candidate compounds based upon the separation of the delta response from a threshold.