Sensor surfaces for detecting analytes

• US 7,208,322 B2
• Filed: 04/02/2002
• Issued: 04/24/2007
• Est. Priority Date: 04/02/2001
• Status: Expired due to Fees

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Stolowitz, Mark L., Li, Guisheng, Lund, Kevin, Wiley, Jean P.
• Primary Examiner(s): Ceperley; Mary E.
• Application Number:
  US10/116,013
• Publication Number:
  US 20020192722A1
• Time in Patent Office: 1,848 Days
• Field of Search: 435/6, 435/7.5, 435/7.2, 436/149, 436/151, 436/806, 436/525, 436/86, 436/528, 436/829, 436/501, 436/94
• US Class Current: 436/525
• 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: G01N 21/553 and using surface plasmons fluorescence excitation G01N21/648; enhanced Raman G01N21/658
• 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
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

The present invention provides a sensor surface comprising: a substrate coated with a free electron metal; and a matrix layer disposed on the free electron metal, wherein the matrix layer comprises an organic compound having a boronic acid complexing moiety. The matrix is preferably a self-assembled monolayer (SAM), a mixed self-assembled monolayer (mSAM), or combinations thereof.

Claims

1. A sensor surface for detecting the interaction of a biological binding pair, said sensor surface comprising:

a substrate coated with a free electron metal;

a matrix layer disposed on said free electron metal, said matrix layer comprising an organic compound, wherein said organic compound has a boronic acid complexing moiety;

a boronic acid moiety complexed to said boronic acid complexing moiety; and

a first member of said binding pair conjugated to said boronic acid moiety, wherein said first member binds to a second member of said binding pair such that a detectable response is elicited.

2. The sensor surface of claim 1, wherein said matrix is a self-assembled monolayer, a mixed self-assembled monolayer, or combinations thereof.

3. The sensor surface of claim 2, wherein said matrix is a mixed self-assembled monolayer.

4. The sensor surface of claim 2, wherein said free electron metal is a member selected from the group consisting of copper, silver, aluminum, platinum and gold.

5. The sensor surface of claim 4, wherein said free electron metal is gold.

6. The sensor surface of claim 1, wherein said organic compound having said boronic acid complexing moiety is of the formula:

X—

R—

Y 

 

(I)

wherein;

X is an anchor group that forms a complex with said free-electron metal;

R is an optionally substituted alkylene group optionally interrupted by one or more members selected from the group consisting of a heteroatom, an amido group and combinations thereof; and

Y is a boronic acid complexing moiety.

7. The sensor surface of claim 6, wherein said organic compound having said boronic acid complexing moiety is of the formula:

X(CH₂)_nC(O)NHR′

-(NSBI)_m-Y 

 

(IIa)

X(CH₂)_nNHC(O)R′

-(NSBI)_m-Y 

 

(IIb)

wherein;

n is an integer from about 1 to 5;

R′

is an optionally substituted alkylene group optionally interrupted by a heteroatom;

NSBI is a nonspecific binding inhibitor;

m is 0 or 1; and

Y is a boronic acid complexing moiety.

8. The sensor surface of claim 7, wherein R′

is an optionally substituted alkylene group, which is optionally interrupted by a heteroatom and which is at least about 8 carbon atoms in length.

9. The sensor surface of claim 7, wherein said nonspecific binding inhibitor is a member selected from the group consisting of an oligo(ethylene glycol), a poly(ethylene glycol), a branched OEG, a branched PEG, an oligo(peptide), an oligo(propylene sulfoxide), a sugar, a sugar alcohol, and a dendrimer.

10. The sensor surface of claim 7, wherein R is an optionally substituted alkylene group, which is optionally interrupted by a heteroatom and which is between about 8 carbon atoms to about 40 carbon atoms in length.

11. The sensor surface of claim 7, wherein m is 1.

12. The sensor surface of claim 7, wherein Y has the formula:

13. The sensor surface of claim 12, wherein said organic compound having said boronic acid complexing moiety is of the formula

HS(CH_2)nC(O)NHR′

-(NSBI)-SHA 

 

(IV)

wherein;

n is an integer from 1 to about 5;

R′

is an optionally substituted alkylene group, which is optionally interrupted by a heteroatom;

NSBI is a nonspecific binding inhibitor;

x is 0 or 1; and

SHA is a salicyihydroxamic acid.

14. The sensor surface of claim 13, wherein R′

is an optionally substituted alkylene group, which is optionally interrupted by a heteroatom and which is at least about 8 carbon atoms in length.

15. The sensor surface of claim 13, wherein said nonspecific binding inhibitor is a member selected from the group consisting of an oligo(ethylene glycol), a poly(ethylene glycol), a branched OEG, a branched PEG, an oligo(peptide), an oligo(propylene sulfoxide), a sugar, a sugar alcohol, and a dendrimer.

16. The sensor surface of claim 13, wherein R′

is an optionally substituted alkylene group, which is optionally interrupted by a heteroatom and which is between about 8 carbon atoms to about 40 carbon atoms in length.

17. The sensor surface of claim 12, wherein said organic compound having said boronic acid complexing moiety is of the formula:

HS(CH₂)₂C(O)NHR′

-(NSBI)-DHBHA 

 

(V)

wherein;

n is an integer from 1 to about 5;

R′

is an optionally substituted alkylene group, which is optionally interrupted by a heteroatom;

NSBI is a nonspecific binding inhibitor; and

DHBHA is a dihydroxybenzohydroxamic acid.

18. The sensor surface of claim 3, wherein said matrix comprises an organic compound of the formula:

X—

(CH₂)_nC(O)NHR′

-(NSBI)_m-Z 

 

(VIa)

or

X—

(CH₂)_nC(O)NHR′

-(NSBI)_m-Z 

 

(VIb)

wherein;

X is an anchor group that forms a complex with a free electron metal;

n is an integer from 1 to 5;

R′

is an optionally substituted alkylene group, which is optionally interrupted by a heteroatom;

NSB is a nonspecific binding inhibitor;

m is 0 or 1; and

Z is an unreactive, uncharged chain-terminating group.

19. The sensor surface of claim 18, wherein said nonspecific binding inhibitor is a member selected from the group consisting of an oligo(ethylene glycol), a poly(ethylene glycol), a branched OEG, a branched PEG, an oligo(peptide), an oligo(propylene sulfoxide), a sugar, a sugar alcohol, and a dendrimer.

20. The sensor surface of claim 18, wherein Z is a member selected from the group consisting of H, CH₃, OH, OCH₃, CO₂CH₃, C(O)NH₂, C(O)NHCH₃, C(O)N(CH₃)₂ or SO₂NH₂.

21. The sensor surface of claim 18, wherein said matrix layer comprises a combination of polymers having the formula:

SCH₂C(O)NH(CH₂)₁₁(OCH₂CH₂)₁₁OC(O)Y (Polymer A), wherein Y is a boronic acid complexing moiety; and

—

SCH₂C(O)NH—

(CH₂)₁₁(COCH₂CH₂)₃OH (Polymer B).

22. The sensor surface of claim 21, wherein the mole percent of Polymer A is less than about 10% of the total mole percent of Polymer A and Polymer B.

23. The sensor surface of claim 21, wherein the mole percent of the Polymer A is less than about 5% of the total mole percent of Polymer A and Polymer B.

24. The sensor surface of claim 18, wherein R′

is an optionally substituted alkylene group, which is optionally interrupted by a heteroatom and which is at least about 8 carbon atoms in length.

25. The sensor surface of claim 18, wherein R′

is an optionally substituted alkylene group, which is optionally interrupted by a heteroatom and which is between about 8 carbon atoms to about 40 carbon atoms in length.

26. The sensor surface of claim 1, wherein said boronic acid moiety comprises a bioactive species.

27. The sensor surface of claim 26, wherein said boronic acid moiety is of the formula:

28. The sensor surface of claim 27 wherein said protein is selected from the group consisting of a capture protein, an antigen, an antibody, protein A, protein G, protein A/G, an enzyme, a lectin, and avidin.

29. The sensor surface of claim 27, wherein said boronic acid moiety has the formula:

30. The sensor surface of claim 27, wherein said boronic acid moiety has the formula:

31. The sensor surface of claim 27, wherein said boronic acid moiety has the formula:

32. The sensor surface of claim 1, wherein said sensor surface is an array of sensor surfaces.

33. The sensor surface of claim 1, wherein said sensor surface is part of a surface plasmon resonance sensing apparatus or a potentiometric sensing apparatus.

34. A sensor surface, said sensor surface comprising:

a substrate coated with a free electron metal;

a matrix layer disposed on said free electron metal, said matrix layer comprising an organic compound, wherein said organic compound has the formula

35. A process for immobilizing a ligand on a sensor surface, said process comprising:

admixing a boronic acid reagent having a reactive group with a ligand to form a boronic acid-ligand conjugate; and

incubating said boronic acid-ligand conjugate with said sensor surface wherein said sensor surface comprises;

a substrate coated with a free electron metal;

a matrix layer disposed on said free electron metal, said matrix layer comprising an organic compound, wherein said organic compound has a boronic acid complexing moiety complexed to the boronic acid-ligand conjugate, thereby immobilizing said ligand.

36. The process for immobilizing a ligand of claim 35, further comprising washing said boronic acid-ligand conjugate complexed to said boronic acid complexing moiety.

37. The process for immobilizing a ligand of claim 35, wherein said sensor surface comprises an array of sensor surfaces.