Process to create biomolecule and/or cellular arrays on metal surfaces and product produced thereby

• US 6,127,129 A
• Filed: 08/05/1999
• Issued: 10/03/2000
• Est. Priority Date: 05/04/1999
• Status: Expired due to Fees

• Current Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Original Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Inventors: Frutos, Anthony G., Corn, Robert M., Brockman, Jennifer M.
• Primary Examiner(s): Campbell, Eggerton A.
• Assistant Examiner(s): LUNDGREN, JEFFREY S
• Application Number:
  US09/368,991
• Time in Patent Office: 425 Days
• Field of Search: 435/6, 430/5
• US Class Current: 435/6.19
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]

Abstract

Disclosed is a process to construct multi-component biomolecule or cellular arrays suitable for use in SPR imaging studies of large molecule, cellular/molecular, and cell/cell interactions. The success of the procedure hinges on the use of a reversible protecting group to modify reversibly ω-functionalized alkanethiols self-assembled on metal substrates.

Claims

1. A method of making a biomolecule or cellular array on a metal substrate comprising:

(a) depositing an ω

-modified alkanethiol monolayer on a metal substrate;

(b) reacting hydrophobic protecting groups with the monolayer;

(c) patterning the monolayer to create an array of exposed metal substrate areas;

(d) depositing ω

-modified alkanethiol in the areas of exposed metal substrate, thereby yielding an array of discrete, unprotected ω

-modified alkanethiol spots;

(e) attaching biomolecules or cells to the discrete, unprotected ω

-modified alkanethiol spots, thereby yielding an array of discrete spots having biomolecules or cells immobilized thereon;

(f) removing the protecting groups of step (b); and

then

(g) making the monolayer resistant to non-specific protein binding wherein the method results in a biomolecule or cellular array.

2. The method of claim 1, wherein in step (a), an amino-C₈ -C₂₄ -alkanethiol is deposited on the metal substrate.

3. The method of claim 1, wherein in step (a), 11-mercaptoundecylamine is deposited on the metal substrate.

4. The method of claim 1, wherein in step (b), 9-fluorenylmethoxycarbonyl-N-hydroxysuccinimide is reacted with the monolayer.

5. The method of claim 1, wherein in step (c), the monolayer is patterned by selective exposure to UV radiation.

6. The method of claim 1, wherein in step (d), 11-mercaptoundecylamine is deposited in the areas of exposed metal substrate.

7. The method of claim 1, wherein in step (e), nucleic acid molecules are attached to the discrete, unprotected ω

-modified alkanethiol spots using a bifunctional linker.

8. The method of claim 1, wherein in step (e), DNA molecules are attached to the discrete, unprotected ω

-modified alkanethiol spots.

9. The method of claim 1, wherein in step (e), RNA molecules are attached to the discrete, unprotected ω

-modified alkanethiol spots.

10. The method of claim 1, wherein in step (e), a heterobifunctional linker is used.

11. The method of claim 10, wherein in step (e), sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate is used as a linker.

12. The method of claim 1, wherein in step (f), the protecting groups are removed by treating the monolayer with a secondary amine.

13. The method of claim 12, wherein in step (f), the protecting groups are removed by treating with a solution selected from the group consisting of tris(2-aminoethyl)amine, ethanolamine, and piperidine.

14. The method of claim 1, wherein in step (g), the monolayer is made resistant to non-specific protein binding by attaching poly(ethylene glycol) moieties thereto.

15. The method of claim 1, wherein in step (a) the ω

-modified alkanethiol monolayer is deposited on a gold substrate.

16. The method of claim 15, wherein in step (a), an amino-C₈ -C₂₄ -alkanethiol is deposited on the gold substrate.

17. The method of claim 15, wherein in step (a), 11-mercaptoundecylamine is deposited on the gold substrate.

18. The method of claim 15, wherein in step (b), 9-fluorenylmethoxycarbonyl-N-hydroxysuccinimide is reacted with the monolayer.

19. The method of claim 15, wherein in step (c), the monolayer is patterned by selective exposure to UV radiation.

20. The method of claim 15, wherein in step (d), 11-mercaptoundecylamine is deposited in the areas of exposed gold substrate.

21. The method of claim 15, wherein in step (e), nucleic acid molecules are attached to the discrete, unprotected ω

-modified alkanethiol spots using a bifunctional linker.

22. The method of claim 15, wherein in step (e), a heterobifunctional linker is used.

23. The method of claim 22, wherein in step (e), sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate is used as a linker.

24. The method of claim 15, wherein in step (f), the protecting groups are removed by treating the monolayer with a secondary amine.

25. The method of claim 15, wherein in step (g), the monolayer is made resistant to non-specific protein binding by attaching poly(ethylene glycol) moieties thereto.