Methods for screening cells and antibodies

• US 8,551,716 B2
• Filed: 06/19/2012
• Issued: 10/08/2013
• Est. Priority Date: 09/09/2002
• Status: Expired due to Term

• Current Assignee: X-Body Incorporated (Bristol-Myers Squibb Company)
• Original Assignee: X-Body Incorporated (Bristol-Myers Squibb Company)
• Inventors: Genick, Christine C., Laing, Lance G., Li, Peter, Smith, Timothy F., Madison, Lara, Lin, Bo
• Primary Examiner(s): Chin, Chris L
• Application Number:
  US13/527,072
• Publication Number:
  US 20130130926A1
• Time in Patent Office: 476 Days
• Field of Search: None
• US Class Current: 435/7.2
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• 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 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/255 Details, e.g. use of specially adapted sources, lighting or optical systems
• CPC Class Codes: G01N 21/4788 Diffraction for sizing particles G01N15/0205
• CPC Class Codes: G01N 21/7743 the reagent-coated grating coupling light in or out of the waveguide
• CPC Class Codes: G01N 33/5026 on cell morphology
• CPC Class Codes: G01N 33/5029 on cell motility
• CPC Class Codes: G01N 33/5032 on intercellular interactions
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G02B 5/1809 with pitch less than or comparable to the wavelength
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays

Abstract

The invention provides methods of detecting a change in cell growth patterns, methods of screening many different antibodies in one receptacle, and methods of detecting specific binding of an antibody to a protein or cell, wherein the antibody is in a mixture of many different antibodies.

Claims

1. A method of screening about 100 or more different antibodies in one receptacle comprising:

(a) immobilizing the antibodies to a colorimetric resonant reflectance biosensor, wherein the colorimetric resonant reflectance biosensor comprises an inner surface of the receptacle;

(b) determining a first peak wavelength value for the receptacle;

(c) adding one or more natively folded proteins to the receptacle;

(d) determining a second peak wavelength value for the receptacle;

(e) comparing the first and second peak wavelength values

wherein, if the second peak wavelength value is higher than the first peak wavelength value, then one or more immobilized antibodies in the receptacle have specifically bound the one or more natively folded proteins.

2. The method of claim 1, wherein the total antibody concentration in the receptacle is greater than 2 mg/ml.

3. The method of claim 1, wherein the one or more antibodies in the receptacle that have specifically bound the one or more natively folded proteins are present at a concentration of less than 5 ng/ml.

4. The method of claim 1, wherein the natively folded protein is a cell surface protein.

5. The method of claim 1, wherein the natively folded protein is part of a whole cell that is added to the receptacle.

6. The method of claim 5, wherein the cell is pre-treated with one or more antibodies prior to the cell being added to the receptacle.

7. The method of claim 1, wherein the antibodies are produced by about 100 or more different hybridoma cells.

8. The method of claim 1, wherein the antibodies are produce by about 1,000 or more different hybridoma cells.

9. A method of detecting specific binding of a first antibody to a protein, wherein the antibody is in a mixture of more than 100 different antibodies, wherein the first antibody is in the mixture of antibodies at a concentration of less than about 3 ng/ml, and wherein the concentration of the mixture of antibodies is greater than about 3 ug/ml comprising:

(a) immobilizing the mixture of antibodies to a colorimetric resonant reflectance biosensor, wherein the colorimetric resonant reflectance biosensor comprises an inner surface of the receptacle;

(b) determining a first peak wavelength value for the receptacle;

(c) adding one or more proteins to the receptacle, wherein one or more of the proteins may specifically bind to the first antibody;

(d) determining a second peak wavelength value for the receptacle;

(e) comparing the first and second peak wavelength values

wherein, if the second peak wavelength value is higher than the first peak wavelength value, then one or more antibodies in the receptacle have specifically bound the one or more of the proteins.

10. A method of detecting specific binding of a first antibody to unpurified cells or unpurified antigen comprising:

(a) immobilizing the first antibody to a colorimetric resonant reflectance biosensor;

(b) detecting a first peak wavelength value;

(c) adding the unpurified cells or unpurified antigen to the colorimetric resonant reflectance biosensor;

(d) detecting a second peak wavelength value;

(e) comparing the first and second peak wavelength values, wherein an increase in the second peak wavelength value indicates specific binding of the first antibody to the unpurified cells or unpurified cells.

11. The method of claim 10, wherein a first antigen specific for the first antibody is added to the unpurified cells or unpurified antigen prior to adding the unpurified cells or unpurified antigen to the colorimetric resonant reflectance biosensor, and wherein a lower second peak wavelength value than the second peak wavelength value of claim 10 indicates specific binding of the unpurified cells or unpurified antigen to the first antigen.

12. The method of claim 10, wherein a second antibody having the same specificity as the first antibody is added to the unpurified cells or unpurified antigen prior to adding the unpurified cells or unpurified antigen to the colorimetric resonant reflectance biosensor, and wherein a lower second peak wavelength value than the second peak wavelength value of claim 10 indicates specific binding of the unpurified cells or unpurified antigen to the second antibody.

13. The method of claim 10, wherein the unpurified cells are about 10,000 or fewer cells.

14. The method of claim 10, wherein the concentration of the first antibody is about 3 ng/ml or less.

15. The method of claim 10, wherein the unpurified cells are present in HAT media, hybridoma media, or cell culture media.

16. The method of claim 10, wherein the first antibody, unpurified cells, and unpurified antigen do not comprise detection labels.