Real time binding analysis of antigens on a biosensor surface
View Patent ↗The invention provides methods for detecting interactions between phage and antigen or antigen and antibody using biosensors.
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.