IP Library Granted Patent US 9,134,307
Granted Patent B2
US 9,134,307 · App. 12/335,433 · Granted Sep 15, 2015

Method for determining ion channel modulating properties of a test reagent

Inventors: Lance G. Laing (Belmont, MA); Rick Wagner (Cambridge, MA); Rafael Fernandez (Jamaica Plain, MA); Alexander Yuzhakov (West Roxbury, MA)
Assignee: X-Body, Inc.
G01N33/54373C12Q1/04G01N33/6872
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Quick Facts
Patent No.
US 9,134,307
App. No.
12/335,433
Granted
Sep 15, 2015
Kind
B2
Abstract

The invention provides methods for identifying modulators of ion channels without the use of recombinant cell lines over-expressing the ion channel proteins or the use of detection labels.

Claims (26)

1. A method of determining ion channel modulating properties of a test reagent comprising:

(a) applying cells to a surface of a colorimetric resonant reflectance optical biosensor;

(b) substantially blocking the functional activity of one or more first types of ion channels, wherein the activity of one or more second types of ion channels is not blocked;

(c) detecting a colorimetric resonant reflectance optical first peak wavelength value (PWV) for the cells;

(d) applying a test reagent to the cells;

(e) detecting a colorimetric resonant reflectance optical second PWV for the cells; and

(f) determining any ion channel modulation properties of the test reagent based on a comparison of the first PWV and second PWV.

2. The method of claim 1 , wherein the functional activity of one or more types of ion channels is substantially blocked by an antibody, protein, or small organic molecule.

3. The method of claim 1 , wherein the one or more first type of ion channels are voltage-gated sodium channels, voltage-gated calcium channels, mechano-sensitive channels, potassium channels, inward-rectifier potassium channels, calcium-activated potassium channels, voltage-gated potassium channels, two pore domain potassium channels, transient receptor potential channels, cation channels of sperm, cyclic nucleotide gated channels, hyperpolaraization activated cyclic nucleotide gated channels, two pore channels, ligand gated channels, and light-gated channels.

4. The method of claim 1 , wherein the one or more second type of ion channels are voltage-gated sodium channels, voltage-gated calcium channels, mechano-sensitive channels, potassium channels, inward-rectifier potassium channels, calcium-activated potassium channels, voltage-gated potassium channels, two pore domain potassium channels, transient receptor potential channels, cation channels of sperm, cyclic nucleotide gated channels, hyperpolaraization activated cyclic nucleotide gated channels, two pore channels, ligand gated channels, and light-gated channels.

5. The method of claim 1 , wherein the cells are incubated for a period of time after: (a) their application to the surface; (b) the blocking the functional activity of one or more first types of ion channels; (c) after the detecting the first PWV for the cells; (d) after the application of the test reagent to the cells; (e) after the detecting the first PWV for the cells, or (f) a combination thereof.

6. The method of claim 1 , wherein the surface of the colorimetric resonant reflectance optical biosensor is an internal surface of a vessel selected from the group consisting of a microtiter well, microtiter plate, test tube, Petri dish, microfluidic channel, and microarray.

7. The method of claim 1 , wherein the first and second PWVs are detected using a scanner with a lens having a lower limit pixel size of about 2 micrometers to about 200 micrometers.

8. The method of claim 1 , wherein the cells and test reagent do not comprise detection labels.

9. The method of claim 1 , wherein the method is performed at a temperature of about 2, 10, 15, 25, 30, or 37 degrees Celsius.

10. The method of claim 1 , further comprising the steps of:

(g) washing the cells;

(h) equilibrating the cells;

(i) optionally repeating steps (a)-(f).

11. The method of claim 1 , wherein the cells are non-recombinant cells.

12. The method of claim 1 , wherein the surface of a colorimetric resonant reflectance optical biosensor is coated with fibronectin prior to the application of the cells to the biosensor.

13. The method of claim 1 , wherein the surface of a colorimetric resonant reflectance optical biosensor is coated with cadherin prior to the application of the cells to the biosensor.

14. The method of claim 1 , wherein the surface of a colorimetric resonant reflectance optical biosensor is coated with collagen prior to the application of the cells to the biosensor.

15. The method of claim 1 , wherein the test reagent is applied to the cells in a potassium-free starvation buffer comprising 150 mM NaCI, 20 mM HEPES, 2 mM calcium chloride; 2 mM magnesium chloride; 5 mM D-(+)-glucose and no potassium.

16. The method of claim 1 , wherein the test reagent is a molecule that potentially modulates the activity of an ion channel or alters the expression of ion channel proteins within cells.

17. The method of claim 1 , wherein the test reagent a molecule that potentially modulates the activity of an ion channel or alters the expression of ion channel proteins within cells selected from the group consisting of a polypeptide, polynucleotide, antibody, small organic molecule, polysaccharide, fatty acid, steroid, purine, pyrimidine, and multivalent cations.

Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 9, 2012
From: SRU BIOSYSTEMS, INC.
To: X-BODY, INC.
Reel/Frame 028014/0817 →
RE-RECORD TO CORRECT THE NAME OF THE FOURTH ASSIGNOR, PREVIOUSLY RECORDED ON REEL 22187 FRAME 0946. Recorded Oct 26, 2009
From: LAING, LANCE; WAGNER, RICK; FERNANDEZ, RAFAEL; YUZHAKOV, ALEXANDER
To: SRU BIOSYSTEMS, INC.
Reel/Frame 023426/0353 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 2, 2009
From: LAING, LANCE; WAGNER, RICK; FERNANDEZ, RAFAEL; YUZAKOV, ALEXANDER
To: SRU BIOSYSTEMS INC
Reel/Frame 022187/0946 →
Continuity (4)
Continuation In Part 12171475 · Jul 11, 2008
Provisional Application 60949142 · Jul 11, 2007
Provisional Application 61043478 · Apr 9, 2008
Related Publication 20090137422A1 · May 28, 2009