System and Method for Performing G Protein Coupled Receptor (GPCR) Cell Assays Using Waveguide-Grating Sensors

• US 20090275074A1
• Filed: 05/20/2009
• Published: 11/05/2009
• Est. Priority Date: 04/05/2005
• Status: Active Grant

• Current Assignee: Corning Incorporated
• Original Assignee: Corning Incorporated
• Inventors: Ferrie, Ann M., Lahiri, Joydeep, Yuen, Po Ki, Fontaine, Norman H., Fang, Ye
• Granted Patent: US 8,148,092 B2
• Time in Patent Office: Days
• US Class Current: 435/29
• CPC Class Codes: G01N 21/4788 Diffraction for sizing particles G01N15/0205
• CPC Class Codes: G01N 21/553 and using surface plasmons fluorescence excitation G01N21/648; enhanced Raman G01N21/658
• CPC Class Codes: G01N 2333/726 G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH
• CPC Class Codes: G01N 2500/10 involving cells
• CPC Class Codes: G01N 33/5041 involving analysis of members of signalling pathways

Abstract

The present invention includes a system and method that uses optical LID biosensors to monitor in real time agonist-induced GPCR signaling events within living cells. Particularly, the present invention includes a system and method for using an optical LID biosensor to screen compounds against a target GPCR within living cells based on the mass redistribution due to agonist-induced GPCR activation. In an extended embodiment, the present invention discloses different ways for self-referencing the optical LID biosensor to eliminate unwanted sensitivity to ambient temperature, pressure fluctuations, and other environmental changes. In yet another extended embodiment, the present invention discloses different ways for screening multiple GPCRs in a single type of cell or multiple GPCRs in multiple types of cells within a single medium solution. In still yet another extended embodiment, the present invention discloses different ways to confirm the physiological or pharmacological effect of a compound against a specific GPCR within living cells.

Claims

1. A method for performing a living cell assay, said method comprising the steps of:

providing a microplate with a well containing a waveguide grating-based biosensor;

placing a cell medium with a first living-cell type within the well to cover the waveguide grating-based biosensor;

culturing the first living-cell type on the surface of the waveguide grating-based biosensor;

applying a solution containing a known agonist or antagonist into the cell medium located on the surface of the waveguide grating-based biosensor;

interrogating the waveguide grating-based biosensor such that an evanescent electromagnetic field extends into the first living-cell type which enables a time dependent response to be obtained, where the time dependent response indicates a mass redistribution response within the first living-cell type; and

analyzing the mass redistribution response to determine if a specific receptor has been expressed in the first living-cell type.

2. A method for performing a living cell assay, said method comprising the steps of:

providing a microplate containing an array of wells, each well incorporating a LID biosensor;

placing a first type of the living cells in a cell medium to cover at least one of the LID biosensors so the first type of the living cells attach to the surface of the at least one LID biosensor;

placing a second type of the living cells in a cell medium to cover at least one of the remaining uncovered LID biosensors so the second type of the living cells attach to the surface of the at least one remaining uncovered LID biosensor;

applying a solution containing a compound into the cell mediums located on the surfaces of the LID biosensors; and

interrogating the LID biosensors to obtain time dependent responses which indicate mass redistribution responses within the first type of living cells and the second type of living cells.

3. The method of claim 2, wherein the interrogating step further includes interrogating the LID biosensors such that evanescent electromagnetic fields extend into the first type of living cells and the second type of living cells which enables the time dependent response to be obtained, where the time dependent responses indicate the mass redistribution responses within the first type of living cells and the second type of living cells.

4. The method of claim 2, wherein the LID biosensor is a waveguide grating-based biosensor.