Molecular recognition at surfaces derivatized with self-assembled monolayers

• US 5,620,850 A
• Filed: 09/26/1994
• Issued: 04/15/1997
• Est. Priority Date: 09/26/1994
• Status: Expired due to Term

• Current Assignee: Harvard University
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Sigal, George B., Whitesides, George M., Bamdad, Cynthia C., Strominger, Jack L.
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): REES, DIANE
• Application Number:
  US08/312,388
• Time in Patent Office: 932 Days
• Field of Search: 435/6, 435/7.1-7.9, 435/4, 536/388.1, 530/300, 252/700, 422/69
• US Class Current: 530/300
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: B82Y 40/00 Manufacture or treatment of nanostructures
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: G01N 2610/00 Assays involving self-assembled monolayers [SAMs]
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/553 Metal or metal coated

Abstract

An article suitable for use as a biosensor includes a species of a formula X--R--Ch adhered to a surface of the article as part of a self-assembled monolayer. X is a functionality that adheres to the surface, R is a spacer moiety, and Ch is a chelating agent. A metal ion can be coordinated by the chelating agent, and a polyamino acid-tagged biological binding partner of a target biological molecule coordinated to the metal ion. A method of the invention involves bringing the article into contact with a medium containing or suspected of containing the target biological molecule and allowing the biological molecule to biologically bind to the binding partner. The article is useful particularly as a surface plasmon resonance chip.

Claims

1. A species having a formula:

space="preserve" listing-type="equation">X--R--Ch--M

wherein X represents a functional group and R represents a spacer moiety that, together, are able to promote formation from solution of a self-assembled monolayer on a surface, Ch represents a chelating agent that coordinates a metal ion, and M represents a metal ion coordinated to the chelating agent, wherein M also is coordinated to a biological binding partner of a biological molecule via coordination sites not filled by the chelating agent or includes coordination sites not filled by the chelating agent that, upon exposure to a polyamino acid tag, are able to become coordinated by the polyamino acid tag.

2. The species as recited in claim 1, wherein the chelating agent is a quadradentate chelating agent.

3. The species as recited in claim 1, wherein the chelating agent is nitrilotriacetic acid.

4. The species as recited in claim 1, wherein the chelating agent and metal ion are selected such that the chelating agent coordinates all but at least two of the metal coordination sites.

5. The species as recited in claim 1, wherein the chelating agent is a tetradentate chelating agent, and the metal ion has a coordination number of six.

6. The species as recited in claim 1, wherein the metal ion is Ni²⁺.

7. The species as recited in claim 1, further comprising a biological binding partner of a biological molecule coordinated to the metal ion.

8. The species as recited in claim 7, wherein the biological binding partner includes a polyamino acid tag that coordinates the metal ion.

9. The species as recited in claim 8, wherein the polyamino acid tag comprises at least two neighboring amino acids defining a chelating agent that coordinates the metal ion.

10. The species as recited in claim 9, wherein the at least two neighboring amino acids are at least two neighboring histidines.

11. The species as recited in claim 7, wherein the biological binding partner specifically binds a protein.

12. A species as recited in claim 1, wherein X represents a functional group that adheres to a gold surface.

13. A species as recited in claim 12, wherein X--R--Ch represents a thiol-linked nitrilotriacetic acid chelate.

14. A species having a formula:

space="preserve" listing-type="equation">X--R--Ch--M

wherein X represents a functional group and R represents a spacer moiety that, together, are able to promote formation from solution of a self-assembled monolayer on a gold surface, Ch represents a chelating agent that coordinates a metal ion, selected from the group consisting of bidentate, tridentate, and quadradentate chelating agents, and M represents a metal ion coordinated to the chelating agent, wherein M also is coordinated to a biological binding partner of a biological molecule via coordination sites not filled by the chelating agent or includes coordination sites not filled by the chelating agent that, upon exposure to a polyamino acid tag, are able to become coordinated by the polyamino acid tag.

15. The species as recited in claim 14, wherein the chelating agent is a quadradentate chelating agent, and the metal ion has a coordination number of six.

16. The species as recited in claim 14, wherein the metal ion is Ni²⁺.

17. The species as recited in claim 14, wherein the chelating agent is nitrilotriacetic acid.

18. A species as recited in claim 14, wherein X--R--Ch represents a thiol-linked nitrilotriacetic acid chelate.

19. The species as recited in claim 14, further comprising a biological binding partner of a biological molecule coordinated to the metal ion.

20. The species as recited in claim 19, wherein the biological binding partner specifically binds a protein.

21. A species having a formula:

space="preserve" listing-type="equation">X--R--Ch--M--BP

wherein X represents a functional group and R represents a spacer moiety that, together, are able to promote formation from solution of a self-assembled monolayer on a surface, Ch represents a chelating agent that coordinates a metal ion, M represents a metal ion coordinated to the chelating agent, and BP represents a biological binding partner of a biological molecule coordinated to the metal ion.

22. The species as recited in claim 21, wherein the biological binding partner includes a polyamino acid tag that coordinates the metal ion.

23. The species as recited in claim 22, wherein the polyamino acid tag comprises at least two neighboring amino acids defining a chelating agent that coordinates the metal ion.

24. The species as recited in claim 23, wherein the at least two neighboring amino acids are at least two neighboring histidines.

25. The species as recited in claim 21, wherein the biological binding partner specifically binds a protein.

26. The species as recited in claim 21, wherein the biological binding partner specifically binds a protein and includes a polyamino acid tag comprising at least two neighboring histidines that coordinates the metal ion.

27. The species as recited in claim 21, wherein X represents a functional group that adheres to a gold surface.

28. A species as recited in claim 21, wherein X--R--Ch represents a thiol-linked nitrilotriacetic acid chelate.