Real time binding analysis of antigens on a biosensor surface

• US 20060148100A1
• Filed: 11/30/2005
• Published: 07/06/2006
• Est. Priority Date: 10/30/2000
• Status: Active Grant

• Current Assignee: X-Body Incorporated (Bristol-Myers Squibb Company)
• Original Assignee: Sru Biosystems Incorporated
• Inventors: Madison, Lara, Gerstenmaier, John
• Granted Patent: US 7,524,625 B2
• Time in Patent Office: Days
• US Class Current: 436/518
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/567 utilising isolate of tissue or organ as binding agent
• CPC Class Codes: G01N 33/6878 in eptitope analysis
• CPC Class Codes: Y10S 436/805 Optical property

Abstract

The invention provides methods for detecting interactions between phage and antigen or antigen and antibody using biosensors.

Claims

1. A method of detecting binding of a binding partner to a phage comprising:

(a) immobilizing a crude phage preparation, unconcentrated phage preparation, non-homogenous phage preparation, concentrated phage preparation or a combination thereof on a biosensor;

(b) contacting the biosensor with the binding partner;

(c) detecting binding of the binding partner to the phage immobilized on the biosensor.

2. The method of claim 1, wherein the binding partner is a small molecule, a carbohydrate, a polymer, a peptide, a soluble protein, a cellular receptor, an antigen mimic of a cellular receptor, a cell, a mammalian cell, or a mammalian cell surface protein.

3. The method of claim 1, wherein the phage preparation and antigen do not comprise a detectable label.

4. The method of claim 1, wherein phage preparation is a phage display library.

5. The method of claim 1, wherein the phage preparation is passively immobilized to the biosensor.

6. The method of claim 1, wherein the phage preparation is immobilized to the biosensor by an antibody specific for a phage coat protein or an antibody fragment specific for a phage coat protein.

7. The method of claim 6, wherein the antibody or antibody fragment is immobilized to the biosensor by binding to a protein that is bound to the biosensor.

8. The method of claim 6, wherein the antibody or antibody fragment comprises a tag.

9. The method of claim 8, wherein the antibody or antibody fragment is immobilized to the biosensor by antibodies specific for the tag.

10. The method of claim 1, wherein the biosensor is a colorimetric resonant reflectance biosensor or an evanescent wave-based biosensor.

11. The method of claim 1, wherein the phage preparation is immobilized in one or more spots on the biosensor surface, in a well, or as one or one more spots in a well.

12. A method for determining epitope classes of antibodies in an antibody population comprising:

(a) immobilizing a display phage, antibody, or antibody fragment to a biosensor;

(b) contacting the biosensor with a binding partner that specifically binds to the display phage, antibody, or antibody fragment immobilized to the biosensor, under conditions suitable for binding of the binding partner to the display phage, antibody, or antibody fragment;

(c) contacting the antibody population with the biosensor;

wherein, a detectable signal generated by binding of the antibody population to the binding partner indicates that different epitope classes are present in the antibody population than in the immobilized display phage, antibody, or antibody fragment.

13. The method of claim 12, wherein the antibody population comprises phage clones.

14. The method of claim 12, wherein the antibody population comprises antibody fragments, full antibodies, phage displaying a full antibody, phage displaying an antibody fragment.

15. The method of claim 12, wherein the antibody population comprises antibodies from a hybridoma.

16. The method of claim 12, wherein the antibody population comprises antibodies from a phage display screen.

17. The method of claim 12, wherein the display phage is a crude phage preparation, an unconcentrated phage preparation, a concentrated phage preparation or a non-homologous phage preparation.

18. The method of claim 12, wherein the binding partner is a small molecule, a carbohydrate, a polymer, a peptide, a soluble protein, a cellular receptor, an antigen mimic of a cellular receptor, a mammalian cell, or a mammalian cell surface protein.

19. The method of claim 18, wherein the mammalian cell surface protein is a membrane-associated protein, a single transmembrane protein, a multi-transmembrane protein, or a protein channel.

20. The method of claim 12, wherein the biosensor is a colorimetric resonant reflectance biosensor or an evanescent wave-based biosensor.

21. The method of claim 12, wherein the antibody population, binding partner and immobilized display phage, immobilized antibody, or immobilized antibody fragment do not comprise a detectable label.