Sensor surfaces for detecting analytes and methods of use

• US 7,179,659 B2
• Filed: 04/02/2002
• Issued: 02/20/2007
• 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:
  US10/115,721
• Publication Number:
  US 20030032202A1
• Time in Patent Office: 1,785 Days
• Field of Search: 435/7.5, 435/6, 435/7.2, 436/149, 436/151, 436/806, 436/525, 436/86, 436/528, 436/829, 436/501, 436/94, 530/410
• 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 method for detecting an analyte, comprising providing a sensor comprising a substrate coated with a free electron metal, a matrix layer disposed on the free electron metal, the matrix layer comprising an organic compound, wherein the organic compound has a boronic acid complexing moiety and, a boronic acid moiety complexed to the boronic acid complexing moiety; and contacting the sensor with the analyte to elicit a response; and measuring the response thereby detecting the analyte.

Claims

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

providing a sensor 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 and, a boronic acid moiety complexed to said boronic acid complexing moiety, wherein said boronic acid moiety is conjugated to a first member of said biological binding pair;

contacting said sensor with a second member of said binding pair, under conditions allowing binding of said second member to said first member of said binding pair, wherein the binding of said second member to said first member elicits a detectable response; and

measuring said response, thereby detecting said interaction of said biological binding pair.

2. The method for detecting the interaction of a biological binding pair of claim 1, wherein said matrix is a self-assembled monolayer.

3. The method for detecting the interaction of a biological binding pair of claim 1, wherein said matrix is a mixed self-assembled monolayer.

4. The method for detecting the interaction of a biological binding pair of claim 1, wherein said free electron metal is a member selected from the group consisting of copper, silver, aluminum, platinum and gold.

5. The method for detecting the interaction of a biological binding pair of claim 4, wherein said free electron metal is gold.

6. The method for detecting the interaction of a biological binding pair 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 method for detecting the interaction of a biological binding pair 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 method for detecting the interaction of a biological binding pair 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 method for detecting the interaction of a biological binding pair of claim 7, wherein R′

0 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.

10. The method for detecting the interaction of a biological binding pair of claim 7, wherein m is 1.

11. The method for detecting the interaction of a biological binding pair of claim 7, wherein Y has the formula:

12. The method for detecting the interaction of a biological binding pair of claim 11, wherein said organic compound having said boronic acid complexing moiety is of the formula

HS(CH₂)_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 salicylhydroxamic acid.

13. The method for detecting the interaction of a biological binding pair of claim 12, wherein R′

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

14. The method for detecting the interaction of a biological binding pair of claim 12, 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.

15. The method for detecting the interaction of a biological binding pair of claim 1, wherein said matrix comprises an organic compound of the formula:

X—

(CH₂)_nC(O)NHR′

—

(NSBI)_m—

Z 

 

(VIa)

X—

(CH₂)_nC(O)NHR′

—

(NSBI)_m—

Z 

 

(VIb)

wherein;

n is an integer from 1 to about 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.

16. The method for detecting the interaction of a biological binding pair of claim 15, 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.

17. The method for detecting the interaction of a biological binding pair of claim 15, 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.

18. The method for detecting the interaction of a biological binding pair of claim 15, wherein said matrix layer comprises a combination of polymers having the formula:

—

SCH₂C(O)NH(CH₂)₁₁(OCH₂CH₂)₁₁OC(O)Y(Polymer A) ; and

—

SCH₂C(O)NH—

(CH₂)₁₁(OCH₂CH₂)₃OH (Polymer B), wherein Y is a boronic acid complexing moiety.

19. The method for detecting the interaction of a biological binding pair of claim 18, wherein the mole percent of Polymer A is less than about 10% of the total mole percent of Polymer A and Polymer B.

20. The method for detecting the interaction of a biological binding pair of claim 18, wherein the mole percent of the Polymer A is less than about 5% of the total mole percent of Polymer A and Polymer B.

21. The method for detecting the interaction of a biological binding pair of claim 15, 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.

22. The method for detecting the interaction of a biological binding pair of claim 15, 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.

23. The method for detecting the interaction of a biological binding pair of claim 1, wherein said second member of said binding pair is bound to said first member of said binding pair.

24. The method for detecting the interaction of a biological binding pair of claim 21, wherein said first member of said biological binding pair comprises a bioactive species.

25. The method for detecting the interaction of a biological binding pair of claim 22, wherein said boronic acid moiety is of the formula:

26. The method for detecting the interaction of a biological binding pair of claim 25, wherein said BAS is a member selected from the group consisting of a protein, a polypeptide, a polypeptide fragment, a nucleic acid, a carbohydrate, a receptor, a hormone, a toxin, a vesicle, a liposome and a cell.

27. The method for detecting the interaction of a biological binding pair of claim 26, wherein said BAS is a protein.

28. The method for detecting the interaction of a biological binding pair 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 AIG, an enzyme, a lectin, and avidin.

29. The method for detecting the interaction of a biological binding pair of claim 25, wherein said boronic acid moiety has the formula:

30. The method for detecting the interaction of a biological binding pair of claim 25, wherein said boronic acid moiety has the formula:

31. The method for detecting the interaction of a biological binding pair of claim 25, wherein said boronic acid moiety has the formula:

32. The method for detecting the interaction of a biological binding pair of claim 1, wherein said sensor surface is an array of sensor surfaces.

33. The method for detecting the interaction of a biological binding pair of claim 1, wherein said sensor surface is used in a surface plasmon resonance sensing or in a potentiometric sensing.