Surface immobilization of biopolymers

• US 5,942,397 A
• Filed: 12/10/1997
• Issued: 08/24/1999
• Est. Priority Date: 12/11/1996
• Status: Expired due to Term

• Current Assignee: Commerce United States of America As Represented By The Secretary of
• Original Assignee: Commerce United States of America As Represented By The Secretary of
• Inventors: Tarlov, Michael J., McKenney, Keith H., Herne, Tonya M.
• Primary Examiner(s): Houtteman, Scott W.
• Application Number:
  US08/988,338
• Time in Patent Office: 622 Days
• Field of Search: 435/6, 536/24.1
• US Class Current: 435/6.12
• 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 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase

Abstract

In one embodiment, the present invention provides a biopolymer-containing monolayer comprising: thiol-derivatized biopolymers and organic thiols bound to a metal substrate. In another embodiment, the present invention provides a method for forming this biopolymer-containing monolayer. Preferably, the biopolymers are single-stranded DNA probes.

Claims

1. A biopolymer-containing monolayer comprising:

a metal substrate;

a plurality of thiol-derivatized oligonucleotides bound to said substrate by a sulfur-substrate linkage, said thiol-derivatized oligonucleotides having the formula;

space="preserve" listing-type="equation">HS--X--Y

where

X is either nothing or an organic linker, and

Y comprises an oligonucleotide terminally bound to X where X is an organic linker or terminally bound to S where X is nothing; and

a plurality of organic thiols bound to said substrate, wherein said plurality of bound thiol-derivatized boligonucleotides and said plurality of bound organic thiols comprise a mixed monolayer.

2. The biopolymer-containing monolayer of claim 1, wherein said thiol-derivatized oligonucleotide comprise thiol-derivatized single-stranded DNA.

3. The biopolymer-containing monolayer of claim 1, wherein said thiol-derivatized oligonucleotides comprise RNA.

4. The biopolymer-containing monolayer of claim 1, wherein said thiol-derivatized oligonucleotides comprise PNA.

5. The biopolymer-containing monolayer of claim 1, wherein Y is a nucleotide sequence of at least six nucleotides.

6. The biopolymer of claim 1, wherein X comprises an organic linker.

7. The biopolymer-containing monolayer of claim 1, wherein said metal substrate comprises a metal selected from the group consisting of the metals gold, silver, platinum, palladium, ruthenium and iridium and alloys of said metals.

8. The biopolymer-containing monolayer of claim 1, wherein said substrate comprises gold.

9. The biopolymer-containing monolayer of claim 1, wherein said organic thiol includes a terminal hydroxy group.

10. The biopolymer-containing monolayer of claim 1, wherein X comprises a hexamethylene linker and said organic thiol compound comprises mercaptohexanol.

11. A method for forming a biopolymer-containing monolayer comprising the steps of:

applying a solution comprising thiol-derivatized oligonucleotides to a substrate comprising a metal to bind said thiol-derivatized oligonucleotides thereto by a sulfur-substrate linkage, the thiol-derivatized oligonucleotides having the formula;

space="preserve" listing-type="equation">HS--X--Y

where

X is either nothing or an organic linker; and

Y comprises an oligonucleotide terminally bound to X where X is an organic linker or terminally bound S where X is nothing; and

applying a solution comprising an organic thiol to displace nonspecifically adsorbed oligonucleotides and to prevent nonspecific adsorption of oligonucleotides.

12. The method of claim 11, further comprising rinsing the substrate to which the thiol-derivatized oligonucleotides and the thiols are bound with water to remove compounds not bound to the substrate.

13. The method of claim 11, wherein said thiol-derivatized oligonucleotides comprise thiol-derivatized single-stranded DNA.

14. The method of claim 11, wherein said thiol-derivatized oligonucleotides comprise RNA.

15. The method of claim 11, wherein said thiol-derivatized oligonucleotides comprise PNA.

16. The method of claim 11, wherein Y is a nucleotide sequence of at least six nucleotides.

17. The method of claim 11, wherein the organic thiol is applied to the substrate by immersing the substrate in an aqueous solution of the organic thiol, thereby removing nonspecifically adsorbed thiol-derivatized oligonucleotides and passivating the surface against nonspecific adsorption of oligonucleotides in solution.

18. The method of claim 11, wherein X comprises an organic linker.

19. The method of claim 11, wherein the substrate comprises a metal selected from the group consisting of the metals gold, silver, platinum, palladium, iridium and ruthenium and alloys of said metals.

20. The method of claim 11, wherein the substrate comprises gold.

21. The method of claim 11, wherein the organic thiol includes a terminal hydroxy group.

22. The method of claim 11, wherein X comprises a hexamethylene linker and said organic thiol comprises mercaptohexanol.