Biosensors for ligand-directed functional selectivity

• US 20090093011A1
• Filed: 11/30/2007
• Published: 04/09/2009
• Est. Priority Date: 10/05/2007
• Status: Abandoned Application

• Current Assignee: Corning Incorporated
• Original Assignee: Corning Incorporated
• Inventors: Fang, Ye, Ferrie, Ann MeeJin
• Application Number:
  US11/998,521
• Publication Number:
  US 20090093011A1
• Time in Patent Office: Days
• US Class Current: 435/29
• CPC Class Codes: C12Q 1/025 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics antimicrobial activity C12Q1/18
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics

Abstract

A system and method for determining ligand-directed functional selectivity of a receptor in a live-cell with a biosensor are disclosed. Also disclosed is a system and method for fragment-based screening with a cell-based, label-free biosensor functional assay.

Claims

1. A method for determining ligand-directed functional selectivity of a receptor of a live-cell, the method comprising:

immobilizing a live-cell having a selected receptor on a biosensor;

contacting the immobilized live-cell with a molecular fragment of a ligand of the receptor;

contacting the molecular fragment contacted immobilized live-cell with a stimulus; and

detecting and comparing the stimulus-induced biosensor response of the live-cell in the presence and absence of the molecular fragment.

2. The method of claim 1 wherein contacting with a molecular fragment and contacting with the stimulus compound is accomplished simultaneously, sequentially, or in reverse order.

3. The method of claim 1 wherein the molecular fragment comprises at least one structural component of a natural ligand of the receptor.

4. The method of claim 1 wherein the molecular fragment is a structural component of a natural agonist for the receptor.

5. The method of claim 4 wherein the receptor is selected from a β

₁-adrenergic receptor, a β

₂-adrenergic receptor, and a combination thereof, and the molecular fragment is selected from catechol, halostachine, or a combination thereof.

6. The method of claim 1 wherein the molecular fragment comprises a partial sequence of a natural ligand if the receptor ligand is a peptide or protein

7. The method of claim 1 wherein the receptor comprises a G protein-coupled receptor (GPCR).

8. The method of claim 7 wherein the G protein-coupled receptor (GPCR) comprises a β

₂-adrenergic receptor (β

₂AR).

9. The method of claim 1 wherein the biosensor response comprises the dynamics, the phases, the signal amplitude, the kinetics of each phase, the transition time from one phase to another phase, or a combination thereof.

10. The method of claim 1 wherein the biosensor response comprises a dynamic mass redistribution if an optical biosensor is selected, a bioimpedance signal if an electrical biosensor is selected, or a combination thereof.

11. The method of claim 1 wherein the stimulus comprises at least one of a natural agonist, a full agonist, strong partial agonist, an antagonist, an inverse agonist, a partial agonist, or a combination thereof.

12. The method of claim 1 wherein the biosensor comprises a label-free apparatus that accomplishes a label-free detection method.

13. The method of claim 1 wherein the molecular fragment modulates cell signaling and the stimulus agonizes, antagonizes, or is independent of the cell signaling response.

14. A method for molecular fragment-based functional screening against a receptor of a live-cell using label-free biosensor, the method comprising:

immobilizing a live-cell having a selected receptor on a biosensor;

contacting the immobilized live-cell with a first molecular fragment;

contacting the immobilized live-cell with a second molecular fragment; and

detecting and comparing the molecular fragment-induced biosensor response of the live-cells.

15. The method of claim 14 wherein contacting with the first molecular fragment and contacting with the second molecular fragment is accomplished simultaneously, sequentially, or in reverse order.