Dual-target biosensor cell assays

• US 8,313,898 B2
• Filed: 02/19/2009
• Issued: 11/20/2012
• Est. Priority Date: 03/05/2008
• Status: Expired due to Fees

• Current Assignee: Corning Incorporated
• Original Assignee: Corning Incorporated
• Inventors: Fang, Ye, Tran, Elizabeth, Randle, David H., Frutos, Anthony Glenn
• Primary Examiner(s): Chong, Kimberly
• Application Number:
  US12/388,685
• Publication Number:
  US 20090226931A1
• Time in Patent Office: 1,370 Days
• Field of Search: 435/6
• US Class Current: 435/6
• 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 2333/71 for growth factors; for growth regulators
• 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 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds
• CPC Class Codes: G01N 33/74 involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors

Abstract

A method and apparatus, as defined herein, for use in compound screening, compound profiling, or both assays, for example, against two different cellular targets in, for example, a single cell-type.

Claims

1. A label-free duplex screening method comprising:

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 at least two different targets;

contacting the immobilized cell with a ligand candidate;

contacting the ligand candidate-treated cell with a mixture containing two markers; and

determining the effect of the ligand candidate on the marker mixture-induced biosensor output.

2. The method of claim 1 wherein the at least two different targets comprise a first target and a second target that are distinguishable from each other by the biosensor.

3. The method of claim 1 wherein each of the markers selectively modulates the activity of one of the different targets.

4. The method of claim 1 wherein the ligand candidate comprises at least one of a chemical compound, a biological molecule, a peptide, a protein, a biological sample, a drug candidate small molecule, a drug candidate biological molecule, or a drug candidate small molecule-biological conjugate.

5. The method of claim 1 wherein the target comprises at least one of a receptor, or a cellular protein.

6. The method of claim 5 wherein the receptor comprises at least one of a G_q-coupled receptor, a G_s-coupled receptor, a G_i-coupled receptor, a G_12/13-coupled receptor, a receptor tyrosine kinase, an ion channel, a sodium-proton exchanger, an integrin receptor, or a transporter.

7. The method of claim 5 wherein the cellular protein comprises at least one of a cellular enzyme, a cellular kinase, or a cellular structural protein.

8. The method of claim 1 wherein the marker comprises at least one of an agonist, a partial agonist, or an inverse agonist, the agonist being capable of activating a target and producing a detectable biosensor output signal.

9. The method of claim 1 wherein the marker comprises at least one of an inhibitor, or an antibody, the marker being capable of activating a target and producing a detectable biosensor output signal.

10. The method of claim 1 wherein each of the markers specifically modulates the activity of a distinct target.

11. The method of claim 10 wherein the targets comprise at least one of a pair of G_q-coupled receptors, a pair of a G_q-coupled receptor and a G_s-coupled receptor, a pair of a G_i-coupled receptor and a G_s-coupled receptor, a pair of a G protein-coupled receptor and a receptor-tyrosine kinase, or a pair of a receptor and a cellular protein.

12. The method of claim 1 wherein the effect of the ligand candidate on the biosensor output comprises modulation of the marker-induced signal responses.

13. The method of claim 12 wherein the modulation comprises a change in signal amplitude, dynamics, kinetics, or a combination thereof.