Real time binding analysis of antigens on a biosensor surface

• US 7,524,625 B2
• Filed: 11/30/2005
• Issued: 04/28/2009
• Est. Priority Date: 10/30/2000
• Status: Expired due to Term

• Current Assignee: X-Body Incorporated (Bristol-Myers Squibb Company)
• Original Assignee: Sru Biosystems Incorporated
• Inventors: Gerstenmaier, John, Madison, Lara
• Primary Examiner(s): Chin; Christopher L
• Application Number:
  US11/290,036
• Publication Number:
  US 20060148100A1
• Time in Patent Office: 1,245 Days
• Field of Search: None
• US Class Current: 435/5
• 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, wherein the crude phage preparation, unconcentrated phage preparation, non-homogenous phage preparation, concentrated phage preparation or a combination thereof is immobilized to the biosensor by an antibody specific for a phage coat protein or an antibody fragment specific for a phage coat protein, and wherein the antibody or antibody fragment comprises a tag;

(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 crude phage preparation, unconcentrated phage preparation, non-homogenous phage preparation, concentrated phage preparation or a combination thereof and binding partner do not comprise a detectable label.

4. The method of claim 1, wherein crude phage preparation, unconcentrated phage preparation, non-homogenous phage preparation, concentrated phage preparation or a combination thereof is a phage display library.

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

6. The method of claim 1, wherein the antibody or antibody fragment is immobilized to the biosensor by binding to a protein that is bound to the biosensor.

7. The method of claim 1, wherein the antibody or antibody fragment is immobilized to the biosensor by antibodies specific for the tag.

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

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

10. The method of claim 1, where the method further comprises determining the quantity of binding partner bound to the to the phage immobilized on the biosensor.

11. 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, wherein the biosensor is a colorimetric resonant reflectance biosensor or an evanescent wave-based biosensor;

(b) contacting the biosensor with the binding partner;

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

12. The method of claim 11, 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.

13. The method of claim 11, wherein the crude phage preparation, unconcentrated phage preparation, non-homogenous phage preparation, concentrated phage preparation or a combination thereof and binding partner do not comprise a detectable label.

14. The method of claim 11, wherein crude phage preparation, unconcentrated phage preparation, non-homogenous phage preparation, concentrated phage preparation or a combination thereof is a phage display library.

15. The method of claim 11, wherein the crude phage preparation, unconcentrated phage preparation, non-homogenous phage preparation, concentrated phage preparation or a combination thereof is passively immobilized to the biosensor.

16. The method of claim 11, wherein the crude phage preparation, unconcentrated phage preparation, non-homogenous phage preparation, concentrated phage preparation or a combination thereof is immobilized to the biosensor by an antibody specific for a phage coat protein or an antibody fragment specific for a phage coat protein.

17. The method of claim 16, wherein the antibody or antibody fragment is immobilized to the biosensor by binding to a protein that is bound to the biosensor.

18. The method of claim 16, wherein the antibody or antibody fragment comprises a tag.

19. The method of claim 18, wherein the antibody or antibody fragment is immobilized to the biosensor by antibodies specific for the tag.

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

21. The method of claim 11, where the method further comprises determining the quantity of binding partner bound to the to the phage immobilized on the bio sensor.

22. 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, wherein the crude phage preparation, unconcentrated phage preparation, non-homogenous phage preparation, concentrated phage preparation or a combination thereof is bound to antibodies specific for a phage coat protein or an antibody fragment specific for a phage coat protein, wherein the antibody or antibody fragment comprises a tag; and

wherein the antibody or antibody fragment is immobilized to the biosensor by antibodies specific for the tag;

(b) contacting the biosensor with the binding partner;

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