Target molecule attachment to surfaces

• US 20030148308A1
• Filed: 07/09/2002
• Published: 08/07/2003
• Est. Priority Date: 09/30/1997
• Status: Active Grant

• Current Assignee: Surmodics Incorporated
• Original Assignee: Surmodics Incorporated
• Inventors: Chappa, Ralph A., Swan, Dale G., Swanson, Melvin J., Guire, Patrick E., Hu, Sheau-Ping
• Granted Patent: US 7,691,787 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00531 essentially square
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00635 by reactive plasma treatment
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00711 Light-directed synthesis
• CPC Class Codes: B01J 2219/00716 Heat activated synthesis
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07B 2200/11 Compounds covalently bound to a solid support
• CPC Class Codes: C07C 235/84 with the carbon atom of at least one of the carboxamide groups bound to a carbon atom of a six-membered aromatic ring
• CPC Class Codes: C07C 323/42 Y being a carbon atom of a six-membered aromatic ring
• CPC Class Codes: C07C 45/63 by introduction of halogen; by substitution of halogen atoms by other halogen atoms
• CPC Class Codes: C07C 49/813 : polycyclic
• 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
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries

Abstract

Method and reagent composition for covalent attachment of target molecules, such as nucleic acids, onto the surface of a substrate. The reagent composition includes groups capable of covalently binding to the target molecule. Optionally, the composition can contain photoreactive groups for use in attaching the reagent composition to the surface. The reagent composition can be used to provide activated slides for use in preparing microarrays of nucleic acids.

Claims

1. A method for preparing an activated support comprising the steps of:

a. providing a support; and

b. coupling a reagent composition to the support, wherein the reagent composition comprises a polymeric backbone having more than one thermochemically amine-reactive or sulfhydryl-reactive groups attached thereto, and wherein the reagent composition is configured and arranged to form covalent bonds with functional groups on a target molecule without use of attracting groups to attract the target molecule to the reagent composition.

2. The method according to claim 1 wherein the support comprises crystalline thermoplastics or amorphous thermoplastics.

3. The method according to claim 1 wherein the reagent composition comprises one or more photoreactive groups.

4. The method according to claim 3 wherein the photoreactive groups are photoreactive aryl ketones selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and heterocyclic analogs of anthrone.

5. The method according to claim 3 wherein the thermochemically reactive groups and photoreactive groups are pendent from the polymeric backbone of the reagent composition.

6. The method according to claim 1 wherein the polymeric backbone of the reagent composition is selected from the group consisting of acrylics, vinyls, nylons, polyurethanes, and polyethers.

7. The method according to claim 1 wherein the thermochemically reactive groups are selected from the group consisting of activated esters, epoxides, azlactones, activated hydroxyls, and maleimide.

8. The method according to claim 1 wherein the step of coupling comprises disposing the reagent composition on the support and treating the reagent composition to couple the reagent composition to the support surface.

9. The method according to claim 3 wherein the step of coupling comprises treating the reagent composition with energy wherein the energy activates the photoreactive groups and covalently couples the reagent composition to the support surface.

10. The method according to claim 1 further comprising the step of using the reagent composition-coupled support for the fabrication of a microarray.

11. A method comprising the steps of:

a. providing a support;

b. disposing a reagent composition on the support, wherein the reagent composition comprises a polymeric backbone having more than one thermochemically amine-reactive or sulfhydryl-reactive groups attached thereto, and wherein the reagent composition is configured and arranged to form covalent bonds with functional groups on a target molecule without use of attracting groups to attract the target molecule to the reagent composition;

c. coupling the reagent composition to the support surface to form a reagent composition-coupled support surface; and

d. disposing at least one target molecule on the reagent composition-coupled support surface, wherein the functional groups of the target molecule forms covalent bonds with the reagent composition.

12. The method according to claim 11 wherein the target molecule is a biomolecule.

13. The method according to claim 12 wherein the biomolecule is a nucleic acid.

14. The method according to claim 13 wherein the nucleic acid comprises one or more functional groups selected from the group consisting of amine and sulfhydryl groups.

15. The method according to claim 11 wherein the reagent composition comprises one or more photoreactive groups.

16. The method according to claim 15 wherein the photoreactive groups are photoreactive aryl ketones selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and heterocyclic analogs of anthrone.

17. The method according to claim 15 wherein the thermochemically reactive groups and photoreactive groups are pendent from the polymeric backbone of the reagent composition.

18. The method according to claim 11 wherein the polymeric backbone of the reagent composition is selected from the group consisting of acrylics, vinyls, nylons, polyurethanes, and polyethers.

19. The method according to claim 11 wherein the thermochemically reactive groups are selected from the group consisting of activated esters, epoxides, azlactones, activated hydroxyls, and maleimide.

20. The method according to claim 11 wherein the support is selected from the group consisting of plastic, silicon hydride, or organosilane-pretreated glass or silicone slides.

21. The method according to claim 11 wherein the support comprises crystalline thermoplastics or amorphous thermoplastics.

22. The method according to claim 11 wherein the step of disposing at least one target molecule is performed with a printing apparatus.

23. The method according to claim 11 wherein the step of disposing at least one target molecule comprises disposing a plurality of different target molecules on the reagent composition-coupled support surface.

24. The method according to claim 23 performed to fabricate a microarray.

25. A method according to claim 24 wherein the microarray includes regions of disposed target molecules which are generally circular in shape, have a diameter of between about 10 microns and about 500 microns, and are separated from other regions in the array by center to center spacing of about 20 microns to about 1000 microns.

26. The method according to claim 11 where, in the step of disposing at least one target molecule, the target molecule is disposed on the reagent composition-coupled support surface in a sample having a volume of twenty nanoliters or less.

27. The method according to claim 11 where, in the step of disposing at least one target molecule, the target molecule is disposed on the reagent composition-coupled support surface in a quantity in the range of 0.1 femtomoles to 10 nanomoles.