Reagent and method for attaching target molecules to a surface

• US 5,858,653 A
• Filed: 09/30/1997
• Issued: 01/12/1999
• Est. Priority Date: 09/30/1997
• Status: Expired due to Term

• Current Assignee: Surmodics Incorporated
• Original Assignee: Surmodics Incorporated
• Inventors: Hu, Sheau-Ping J., Duran, Lise W., Swanson, Melvin J., Guire, Patrick E., Amos, Richard A.
• Primary Examiner(s): Richter, Johann
• Assistant Examiner(s): Oswecki, Jane C.
• Application Number:
  US08/940,213
• Time in Patent Office: 469 Days
• Field of Search: 435/6, 435/176, 435/180, 436/501, 436/96, 436/112, 436/120, 436/128, 436/905, 548/542, 560/106, 564/155, 568/332
• US Class Current: 435/6.12
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• 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 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00711 Light-directed synthesis
• CPC Class Codes: B01J 2219/00716 Heat activated synthesis
• 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: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: Y10S 436/905 Photochemical activation of reactions
• CPC Class Codes: Y10T 436/145555 Hetero-N
• CPC Class Codes: Y10T 436/174614 Tertiary amine
• CPC Class Codes: Y10T 436/182 : Organic or sulfhydryl containing [e.g., mercaptan, hydrogen, sulfide, etc.]
• CPC Class Codes: Y10T 436/200833 : Carbonyl, ether, aldehyde or ketone containing

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 attracting the target molecule as well as groups capable of covalently binding to the target molecule, once attracted. Optionally, the composition can contain photoreactive groups for use in attaching the reagent composition to the surface.

Claims

1. A reagent composition for attaching a target molecule to the surface of a substrate, the reagent composition comprising one or more groups for attracting the target molecule to the reagent, and one or more thermochemically amine-reactive or sulfhydryl-reactive groups for forming covalent bonds with corresponding functional groups on the attracted target molecule.

2. A reagent composition according to claim 1 wherein the reagent composition further comprises one or more photoreactive groups for attaching the reagent composition to the surface upon application of energy from a source of electromagnetic radiation.

3. A reagent composition according to claim 1 wherein the attracting groups and thermochemically reactive groups are pendent upon one or more hydrophilic polymeric backbones.

4. A reagent composition according to claim 1 wherein the target molecule is a nucleic acid.

5. A reagent composition according to claim 1 wherein the attracting groups are ionic groups.

6. A reagent composition according to claim 5 wherein the ionic groups are selected from the group consisting of quaternary ammonium groups and protonated tertiary amines.

7. A reagent composition according to claim 4 wherein the nucleic acid comprises a functional group selected from the group consisting of amine and sulfhydryl groups.

8. A reagent composition according to claim 4 wherein the reagent composition comprises a hydrophilic polymeric backbone comprising one or more ionic groups as attracting groups and one or more photoreactive groups for attaching the reagent composition to the surface upon application of energy from a source of electromagnetic radiation.

9. A reagent composition according to claim 8 wherein the ionic groups comprise quaternary ammonium groups.

10. A reagent composition according to claim 2 wherein the photoreactive groups are selected from the group consisting of photoreactive aryl ketones.

11. A reagent composition according to claim 10 wherein the photoreactive aryl ketones are each, independently, selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and anthrone-like heterocycles.

12. A reagent composition according to claim 2 wherein the target molecule is a nucleic acid, the attracting groups are ionic groups, and the photoreactive groups are selected from the group consisting of photoreactive aryl ketones.

13. A reagent composition according to claim 12 wherein the photoreactive aryl ketones are each, independently, selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and anthrone-like heterocycles.

14. A reagent composition according to claim 3 wherein the reagent composition is provided in the form of a composition comprising a first reagent component comprising a hydrophilic backbone comprising one or more attractive groups and one or more photoreactive groups, and a second reagent component comprising a hydrophilic backbone comprising one or more thermochemically amine-reactive or sulfhydryl-reactive groups and one or more photoreactive groups.

15. A reagent composition according to claim 14 wherein the target molecule is a nucleic acid.

16. A reagent composition according to claim 14 wherein the attracting groups are ionic groups.

17. A reagent composition according to claim 16 wherein the ionic groups are selected from the group consisting of quaternary ammonium groups and protonated tertiary amines.

18. A reagent composition according to claim 15 wherein the nucleic acid comprises a functional group selected from the group consisting of amine and sulfhydryl groups.

19. A reagent composition according to claim 14 wherein the photoreactive groups are selected from the group consisting of photoreactive aryl ketones.

20. A reagent composition according to claim 19 wherein the photoreactive aryl ketones are each, independently, selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and anthrone-like heterocycles.

51. A surface comprising a coated reagent composition of claim 1.

52. A surface according to claim 51 wherein the reagent composition further comprises one or more photoreactive groups for attaching the reagent composition to the surface upon application of energy from a source of electromagnetic radiation.

53. A surface according to claim 52 wherein the target molecule is a nucleic acid.

54. A surface according to claim 51 wherein the attracting groups are ionic groups selected from the group consisting of quaternary ammonium groups and protonated tertiary amines.

55. A surface according to claim 53 wherein the nucleic acid comprises a functional group selected from the group consisting of amine and sulfhydryl groups.

56. A surface according to claim 51 wherein the photoreactive groups are selected from the group consisting of photoreactive aryl ketones.

57. A surface according to claim 56 wherein the photoreactive aryl ketones are each, independently, selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and anthrone-like heterocycles.

58. A surface according to claim 52 wherein the reagent composition is provided in the form of a composition comprising a first reagent component comprising a hydrophilic backbone comprising one or more attractive groups and one or more photoreactive groups, and a second reagent component comprising a hydrophilic backbone comprising one or more thermochemically amine-reactive or sulfhydryl-reactive groups and one or more photoreactive groups.

59. A surface according to claim 58 wherein the photoreactive groups are selected from the group consisting of photoreactive aryl ketones.

60. A surface according to claim 59 wherein the photoreactive aryl ketones are each, independently, selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and anthrone-like heterocycles.

21. A method for attaching a target molecule to the surface of a substrate, the method comprising the steps of (a) providing upon the surface of the substrate a reagent composition comprising one or more groups for attracting the target molecule to the reagent composition, and one or more thermochemically amine-reactive or sulfhydryl-reactive groups for forming covalent bonds with corresponding functional groups on the attracted target molecule, (b) bringing the target molecule into sufficient proximity to the surface to permit the attractive groups to attract the target molecule to the bound reagent composition, and (c) allowing the thermochemically reactive groups to form covalent bonds with the attracted target molecule.

22. A method according to claim 21 wherein the reagent composition further comprises one or more photoreactive groups for attaching the reagent composition to the surface upon application of energy from a source of electromagnetic radiation.

23. A method according to claim 21 wherein the attracting groups and thermochemically reactive groups are pendent upon one or more hydrophilic polymeric backbones.

24. A method according to claim 21 wherein the target molecule is a nucleic acid.

25. A method according to claim 21 wherein the attracting groups are ionic groups.

26. A method according to claim 25 wherein the ionic groups are selected from the group consisting of quaternary ammonium groups and protonated tertiary amines.

27. A method according to claim 24 wherein the nucleic acid comprises a functional group selected from the group consisting of amine and sulfhydryl groups.

28. A method according to claim 24 wherein the reagent composition comprises a hydrophilic polymeric backbone comprising one or more ionic groups as attracting groups and one or more photoreactive groups for attaching the reagent composition to the surface upon application of energy from a source of electromagnetic radiation.

29. A method according to claim 28 wherein the ionic groups comprise quaternary ammonium groups.

30. A method according to claim 22 wherein the photoreactive groups are selected from the group consisting of photoreactive aryl ketones.

31. A method according to claim 30 wherein the photoreactive aryl ketones are each, independently, selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and anthrone-like heterocycles.

32. A method according to claim 26 wherein the target molecule is a nucleic acid and the photoreactive groups are selected from the group consisting of photoreactive aryl ketones.

33. A method according to claim 32 wherein the photoreactive aryl ketones are each, independently, selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and anthrone-like heterocycles.

34. A method according to claim 23 wherein the reagent composition is provided in the form of a composition comprising a first reagent component comprising a hydrophilic backbone comprising one or more attractive groups and one or more photoreactive groups, and a second reagent component comprising a hydrophilic backbone comprising one or more thermochemically amine-reactive or sulfhydryl-reactive groups and one or more photoreactive groups.

35. A method according to claim 34 wherein the target molecule is a nucleic acid.

36. A method according to claim 31 wherein the attracting groups are ionic groups.

37. A method according to claim 36 wherein the ionic groups are selected from the group consisting of quaternary ammonium groups and protonated tertiary amines.

38. A method according to claim 35 wherein the nucleic acid comprises a functional group selected from the group consisting of amine and sulfhydryl groups.

39. A method according to claim 34 wherein the photoreactive groups are selected from the group consisting of photoreactive aryl ketones.

40. A method according to claim 39 wherein the photoreactive aryl ketones are each, independently, selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and anthrone-like heterocycles.

41. A substrate surface coated with a target molecule and a reagent composition by the method of claim 21.

42. A surface according to claim 41 wherein the reagent composition further comprises one or more photoreactive groups for attaching the reagent composition to the surface upon application of energy from a source of electromagnetic radiation.

43. A surface according to claim 42 wherein the target molecule is a nucleic acid.

44. A surface according to claim 41 wherein the attracting groups are ionic groups selected from the group consisting of quaternary ammonium groups and protonated tertiary amines.

45. A surface according to claim 43 wherein the nucleic acid comprises a functional group selected from the group consisting of amine and sulfhydryl groups.

46. A surface according to claim 41 wherein the photoreactive groups are selected from the group consisting of photoreactive aryl ketones.

47. A surface according to claim 46 wherein the photoreactive aryl ketones are each, independently, selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and anthrone-like heterocycles.

48. A surface according to claim 42 wherein the reagent composition is provided in the form of a composition comprising a first reagent component comprising a hydrophilic backbone comprising one or more attractive groups and one or more photoreactive groups, and a second reagent component comprising a hydrophilic backbone comprising one or more thermochemically amine-reactive or sulfhydryl-reactive groups and one or more photoreactive groups.

49. A surface according to claim 48 wherein the photoreactive groups are selected from the group consisting of photoreactive aryl ketones.

50. A surface according to claim 49 wherein the photoreactive aryl ketones are each, independently, selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and anthrone-like heterocycles.