Methods of detection of changes in cells

• US 8,298,780 B2
• Filed: 12/15/2008
• Issued: 10/30/2012
• Est. Priority Date: 09/22/2003
• Status: Active Grant

• Current Assignee: X-Body Incorporated (Bristol-Myers Squibb Company)
• Original Assignee: X-Body Incorporated (Bristol-Myers Squibb Company)
• Inventors: Wagner, Rick, Fernandez, Rafael, Cunningham, Brian T., Laing, Lance
• Primary Examiner(s): Chin, Chris L
• Application Number:
  US12/335,393
• Publication Number:
  US 20090130703A1
• Time in Patent Office: 1,415 Days
• Field of Search: None
• US Class Current: 435/7.2
• CPC Class Codes: C40B 30/04 by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
• CPC Class Codes: C40B 30/10 by measuring physical properties, e.g. mass
• CPC Class Codes: G01N 21/4788 Diffraction for sizing particles G01N15/0205
• CPC Class Codes: G01N 21/63 optically excited
• CPC Class Codes: G01N 21/7743 the reagent-coated grating coupling light in or out of the waveguide
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings

Abstract

Methods are provided to detect changes in cells without the use of detection labels.

Claims

1. A method of detection of responses of cells to stimuli comprising:

(a) (i) immobilizing one or more extracellular matrix ligands to a surface of a colorimetric resonant reflectance biosensor or a grating-based waveguide biosensor, wherein the cells have cell surface receptors specific for the one or more extracellular matrix ligands; and

adding the cells to the biosensor;

or

(ii) mixing cells with one or more extracellular matrix ligands, wherein the cells have cell surface receptors specific for the one or more extracellular matrix ligands; and

adding the cells with one or more extracellular matrix ligands to a surface of a colorimetric resonant reflectance biosensor or a grating-based waveguide biosensor;

(b) exposing the cells to stimuli, wherein the stimuli are compounds that modulate activity of a G protein-coupled receptor, ion channels, P13 kinase, transient receptor potential channels, phospholipase C, receptor tyrosine kinases, cytokines, beta-arrestin pathway responses, cytoskeletal rearrangements, epigenetic signals, integrins, or signal transduction pathways; and

(c) detecting a change in peak wavelength value or signal over time, which indicates that the cells have responded to the stimuli.

2. The method of claim 1, wherein the cells of (a) are in serum-free medium.

3. The method of claim 2, wherein the exposing of the cells to stimuli is done without washing off the serum-free medium.

4. The method of claim 1, wherein prior to (a) the cells are washed with buffered saline.

5. The method of claim 4, wherein the buffered saline substantially lacks calcium and magnesium.

6. The method of claim 1, wherein prior to step (a), the following steps are performed:

(i) washing the cells with a buffered saline solution lacking calcium and magnesium;

(ii) removing the buffered saline solution lacking calcium and magnesium from the cells;

(iii) adding an isotonic ethylenediaminetetraacetate chealating agent to the cells;

(iv) neutralizing the isotonic ethylenediaminetetraacetate chealating agent with buffered saline solution.

7. The method of claim 1, wherein the cells are added to the biosensor surface in the presence of an adhesion modulator.

8. The method of claim 1, wherein the detecting the response of the cells to the stimuli is done with a detection device with a resolution of about 2 to 10 micrometers.

9. The method of claim 1, wherein the response of the cells to stimuli are changes in cell growth patterns, changes in cell death patterns, changes in cell movement, changes in cell size or volume, or changes in cell adhesion.