Production of polymeric microarrays

• US 20040028804A1
• Filed: 08/07/2002
• Published: 02/12/2004
• Est. Priority Date: 08/07/2002
• Status: Abandoned Application

• Current Assignee: Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Anderson, Daniel G., Langer, Robert
• Application Number:
  US10/214,723
• Publication Number:
  US 20040028804A1
• Time in Patent Office: Days
• US Class Current: 427/2.11
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00387 Applications using probes
• CPC Class Codes: B01J 2219/00497 Features relating to the solid phase supports
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• 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 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00628 Ionic
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00675 In-situ synthesis on the substrate
• CPC Class Codes: B01J 2219/00691 using robots
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• 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: B01J 2219/00725 : Peptides
• CPC Class Codes: B01J 2219/00736 : Non-biologic macromolecules, e.g. polymeric compounds
• CPC Class Codes: B01J 2219/00743 : Cells
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 40/14 : Libraries containing macromolecular compounds and not covered by groups C40B40/06 - C40B40/12
• CPC Class Codes: C40B 50/14 : Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 2650/00 : Assays involving polymers whose constituent monomers bore biological functional groups before polymerization, i.e. vinyl, acryl derivatives of amino acids, sugars

Abstract

A method of preparing a microarray of polymer elements. The method includes providing a substrate surface, providing a plurality of individual monomers in a liquid phase, depositing said monomers as a plurality of discrete monomer elements on the substrate surface, and exposing the plurality of discrete monomer elements to initiating conditions to form polymer elements. A portion of the discrete monomer elements may include more than one type of monomer.

Claims

1. A method of preparing a microarray of polymer elements comprising:

providing a substrate surface;

providing a plurality of individual monomers in a liquid phase;

depositing said monomers as a plurality of discrete monomer elements on said substrate surface; and

exposing the plurality of discrete monomer elements to initiating conditions so that polymer elements are formed.

2. The method of claim 1, wherein at least a portion of the discrete elements include more than one type of monomer.

3. The method of claim 1, wherein at least a portion of the monomers are a liquid at room temperature.

4. The method of claim 3, wherein the liquid is combined with a solvent.

5. The method of claim 1, wherein at least a portion of the monomers are a solid at room temperature and providing the monomers in a liquid phase comprises dissolving the monomers in a solvent at a concentration of about 3M or less.

6. The method of claim 1, wherein the polymer elements are a first portion of the microarray of polymer elements, and the method further comprises repeating the steps of depositing and exposing to prepare a second portion of the array of polymer elements.

7. The method of claim 1, wherein the steps of depositing and exposing comprise:

A) depositing a single discrete monomer element on said substrate surface using a robotic liquid handling device;

B) exposing the discrete monomer element to initiating conditions; and

repeating steps a) and b) until a predetermined number of polymer elements have been prepared.

8. The method of claim 1, wherein the substrate comprises a material selected from the group consisting of glass, plastic, metal, ceramic, and combinations thereof.

9. The method of claim 1, wherein the step of providing a substrate surface comprises modifying a surface chemistry of the substrate.

10. The method of claim 9, wherein modifying a surface chemistry comprises a member of the group consisting of introducing crystallographic texture, oxidizing, sulfidating, patterning, covalently attaching a functional group, and any combination of the above.

11. The method of claim 9, wherein modifying a surface chemistry comprises depositing a polymer on the surface.

12. The method of claim 11, wherein the polymer is cytophobic.

13. The method of claim 11, wherein the polymer is a hydrogel.

14. The method of claim 1, wherein the polymer elements are bound to the substrate surface via an interaction selected from the group consisting of covalent interactions, chemical adsorption, hydrogen bonding, surface interpenetration, ionic bonding, van der Waals forces, hydrophobic interactions, magnetic interactions, dipole-dipole interactions, mechanical interlocking, and combinations of these.

15. The method of claim 1, wherein the polymer elements are non-biocompatible.

16. The method of claim 1, wherein the polymer elements are biocompatible.

17. The method of claim 16, wherein the polymer elements include monomers of polymers selected from polyamides, polyphosphazenes, polypropylfumarates, synthetic poly(amino acids), polyethers, polyacetals, polycyanoacrylates, polyurethanes, polycarbonates, polyanhydrides, poly(ortho esters), polyhydroxyacids, polyesters, polyacrylates, ethylene-vinyl acetate polymers, cellulose acetates, polystyrenes, chlorosulphonated polyolefins, polyaniline, polyesters, polyamides, polymerized vinyl compounds, and polymerized vinylidine compounds.

18. The method of claim 1, further comprising including a compound selected from the group consisting of drugs, growth factors, combinatorial compounds, proteins, polysaccharides, polynucleotides, and lipids in at least a portion of the polymer elements.

19. The method of claim 18, wherein the compound is covalently attached to at least a portion of the polymer elements.

20. The method of claim 19, wherein the compound is functionalized with a moiety that is incorporated into the polymer element during polymerization, and wherein the step of including comprises depositing the functionalized compound on at least one predetermined discrete monomer element.

21. The method of claim 20, wherein the moiety is a member of an acrylate group, a vinyl group, an acrylamide, and an epoxide.

22. The method of claim 20, wherein the moiety includes a photoreactive chemical group that initiates polymerization upon exposure to UV light.

23. The method of claim 19, wherein the compound is incorporated into a backbone of the polymer of the polymer element.

24. The method of claim 18, wherein the compound is non-covalently bound to the polymer of the polymer element.

25. The method of claim 1, wherein the polymer elements are spaced at intervals between about 300 μ

m and about 1200 μ

m.

26. The method of claim 1, wherein the polymer elements are spaced at intervals of less than about 300 μ

m.

27. The method of claim 1, wherein the polymer elements are spaced at intervals of less than about 1 μ

m.

28. The method of claim 1, wherein the polymer elements are spaced at intervals of less than about 0.1 μ

m.

29. The method of claim 1, further comprising seeding cells on the polymer elements.

30. The method of claim 29, wherein the cells are selected from the group consisting of yeast cells, mammalian cells, bacterial cells, and plant cells.

31. The method of claim 30, wherein said cells are selected from the group of mammalian cells consisting of chondrocytes, fibroblasts, connective tissue cells, epithelial cells, endothelial cells, cancer cells, hepatocytes, islet cells, smooth muscle cells, skeletal muscle cells, heart muscle cells, kidney cells, intestinal cells, organ cells, lymphocytes, blood vessel cells, stem cells, human embryonic stem cells, and mesenchymal stem cells.

32. The method of claim 1, wherein the step of depositing is performed with a robotic liquid handling device.

33. The method of claim 32, wherein the liquid handling device deposits via a member of the group consisting of pin fluid deposition, syringe pumped fluid deposition, and piezoelectric fluid deposition.

34. The method of claim 1, wherein the monomers are deposited as drops of between about 0.1 and about 100 nL.

35. The method of claim 34, wherein the monomers are deposited as drops of between 1 and 10 nL.

36. The method of claim 1, wherein the monomers are deposited as drops of less than about 0.1 nL.

37. The method of claim 1, wherein the initiating conditions are selected from the group consisting of exposure to UV light, an increase in temperature, exposure to an environment containing water vapor, exposure to an environment containing oxygen, and any combination of the above.

38. The method of claim 1, further comprising depositing a chemical initiator on the discrete monomer elements, wherein the chemical initiator is co-deposited with at least a portion of the monomers, deposited separately from at least a portion of the monomers, or co-deposited with a first portion of the monomers and deposited on the discrete elements separately from a second portion of the monomers.

39. The method of claim 38, wherein the initiator is selected from a radical initiator, a redox initiator, a thermal initiator, and an ionic initiator.

40. The method of claim 1, wherein the monomers are selected from the group consisting of 1,4 butanediol dimethacrylate, diethylene glycol diacrylate, diethylene glycol dimethacrylate, 1,6 hexanediol diacrylate, neopentyl glycol diacrylate, phenylene diacrylate 1,3, propoxylated neopentyl glycol diacrylate, tetraethylene glycol diacrylate, tetraethylene glycol dimethacrylate, triethylene glycol diacrylate, triethylene glycol dimethacrylate, tripropylene glycol diacrylate, caprolactone 2-(methacryloyloxy)ethyl ester, 5-ethyl-5-(hydroxymethyl)-β

,β

-dimethyl-1,3-dioxane-2-ethanol diacrylate, 1,6-hexanediol propoxylate diacrylate, 3-hydroxy-2,2-dimethylpropyl 3-hydroxy-2,2-dimethylpropionate diacrylate, glycerol 1,3-diglycerolate diacrylate, glycerol dimethacrylate, mixture of isomers, tech., 85%, neopentyl glycol dimethacrylate, neopentyl glycol ethoxylate (1 EO/OH) diacrylate, trimethylolpropane benzoate diacrylate, 1,14-tetradecanediol dimethacrylate, tricyclo[5.2.1.0^2,6]decanedimethanol diacrylate, trimethylolpropane ethoxylate (1 EO/OH) methyl ether diacrylate, and trimethylolpropane triacrylate, tech.

41. A microarray of polymers comprising a plurality of discrete polymer elements bound to a surface, said microarray being produced by the steps of:

providing solutions of monomers of polymer materials in a liquid phase;

depositing at least a first portion of said monomers as a plurality of discrete monomer elements on said surface; and

depositing at least a second portion of said monomers on a portion of said discrete monomer elements; and

exposing the plurality of discrete monomer elements to initiating conditions so that polymer elements are formed.

42. The microarray of claim 41, wherein a portion of said solutions include a biomolecule derivatized with a moiety that is covalently incorporated into the polymer element after the step of exposing.

43. The microarray of claim 42, wherein the moiety is selected from the group consisting of an acrylate group, a vinyl group, an acrylamide, and an epoxide.

44. The microarray of claim 42, wherein the moiety includes a photoreactive chemical structure that initiates polymerization after the step of exposing.

45. The microarray of claim 41, wherein at least a portion of the monomers are a liquid at room temperature.

46. The microarray of claim 45, wherein the liquid is combined with a solvent.

47. The microarray of claim 41, wherein at least a portion of the monomers are a solid at room temperature and providing the monomers a liquid phase comprises dissolving the monomers in a solvent at a concentration of about 3M or less.

48. The microarray of claim 41, wherein the plurality of discrete polymer elements are a first portion of the microarray of polymer elements, and wherein the steps of depositing and exposing are repeated to prepare a second portion of the array of polymer elements.

49. The microarray of claim 41, wherein each discrete monomer element is deposited and exposed individually.

50. The microarray of claim 41, wherein the surface comprises a material selected from the group consisting of glass, polymer, metal, ceramic, and combinations thereof.

51. The microarray of claim 50, wherein the surface is cytophobic.

52. The microarray of claim 50, wherein the surface is a hydrogel.

53. The microarray of claim 41, wherein a chemistry of the surface is modified.

54. The microarray of claim 53, wherein the chemistry of the surface is modified by a member of the group consisting of introducing crystallographic texture, oxidizing, sulfidating, patterning, covalently attaching a functional group, and any combination of the above.

55. The microarray of claim 41, wherein the polymer elements are bound to the surface via an interaction selected from the group consisting of covalent interactions, chemical adsorption, hydrogen bonding, surface interpenetration, ionic bonding, van der Waals forces, hydrophobic interactions, magnetic interactions, dipole-dipole interactions, mechanical interlocking, and combinations of these.

56. The microarray of claim 41, wherein the polymer elements are non-biocompatible.

57. The microarray of claim 41, wherein the polymer elements are biocompatible.

58. The microarray of claim 57, wherein the polymer elements include monomers of polymers selected from polyamides, polyphosphazenes, polypropylfumarates, synthetic poly(amino acids), polyethers, polyacetals, polycyanoacrylates, polyurethanes, polycarbonates, polyanhydrides, poly(ortho esters), polyhydroxyacids, polyesters, polyacrylates, ethylene-vinyl acetate polymers, cellulose acetates, polystyrenes, chlorosulphonated polyolefins, polyaniline, polyesters, polyamides, polymerized vinyl compounds, and polymerized vinylidine compounds.

59. The microarray of claim 41, further comprising including a compound selected from the group consisting of drugs, growth factors, combinatorial compounds, proteins, polysaccharides, polynucleotides, and lipids in at least a portion of the polymer elements.

60. The microarray of claim 59, wherein the compound is covalently attached to at least a portion of the polymer elements.

61. The microarray of claim 60, wherein the compound is functionalized with a moiety that is incorporated into the polymer element during polymerization, and wherein the functionalized compound is deposited on at least one predetermined discrete monomer element.

62. The microarray of claim 61, wherein the moiety is a member of an acrylate group, a vinyl group, an acrylamide, and an epoxide.

63. The microarray of claim 61, wherein the moiety includes a photoreactive chemical group that initiates polymerization upon exposure to UV light.

64. The microarray of claim 60, wherein the compound is incorporated into a backbone of the polymer of the polymer element.

65. The microarray of claim 59, wherein the compound is non-covalently bound to the polymer of the polymer element.

66. The microarray of claim 41, wherein the polymer elements are spaced at intervals between about 300 μ

m and about 1200 μ

m.

67. The microarray of claim 41, wherein the polymer elements are spaced at intervals of less than about 300 μ

m.

68. The microarray of claim 41, wherein the polymer elements are spaced at intervals of less than about 1 μ

m.

69. The microarray of claim 41, wherein the polymer elements are spaced at intervals of less than about 0.1 μ

m.

70. The microarray of claim 41, wherein cells are seeded on the polymer elements.

71. The microarray of claim 70, wherein the cells are selected from the group consisting of yeast cells, mammalian cells, bacterial cells, and plant cells.

72. The microarray of claim 71, wherein said cells are selected from the group of mammalian cells consisting of chondrocytes, fibroblasts, connective tissue cells, epithelial cells, endothelial cells, cancer cells, hepatocytes, islet cells, smooth muscle cells, skeletal muscle cells, heart muscle cells, kidney cells, intestinal cells, organ cells, lymphocytes, blood vessel cells, stem cells, human embryonic stem cells, and mesenchymal stem cells.

73. The microarray of claim 41, wherein the steps of depositing are performed with a robotic liquid handling device.

74. The microarray of claim 73, wherein the robotic liquid handling device deposits via a member of the group consisting of pin fluid deposition, syringe pumped fluid deposition, and piezoelectric fluid deposition.

75. The microarray of claim 41, wherein the monomers are deposited as drops of between about 0.1 and about 100 nL.

76. The microarray of claim 75, wherein the monomers are deposited as drops of between 1 and 10 nL.

77. The microarray of claim 41, wherein the monomers are deposited as drops of less than about 0.1 nL.

78. The microarray of claim 41, wherein the initiating conditions are selected from the group consisting of exposure to UV light, an increase in temperature, exposure to an environment containing water vapor, exposure to an environment containing oxygen, and any combination of the above.

79. The microarray of claim 41, wherein a chemical initiator is deposited on the discrete monomer elements, wherein the chemical initiator is co-deposited with at least a portion of the monomers, deposited separately from at least a portion of the monomers, or co-deposited with the first portion of the monomers and deposited on the discrete elements separately from the second portion of the monomers.

80. The microarray of claim 79, wherein the initiator is selected from a radical initiator, a redox initiator, a thermal initiator, and an ionic initiator.

81. The microarray of claim 41, wherein the monomers are selected from the group consisting of 1,4 butanediol dimethacrylate, diethylene glycol diacrylate, diethylene glycol dimethacrylate, 1,6 hexanediol diacrylate, neopentyl glycol diacrylate, phenylene diacrylate 1,3, propoxylated neopentyl glycol diacrylate, tetraethylene glycol diacrylate, tetraethylene glycol dimethacrylate, triethylene glycol diacrylate, triethylene glycol dimethacrylate, tripropylene glycol diacrylate, caprolactone 2-(methacryloyloxy)ethyl ester, 5-ethyl-5-(hydroxymethyl)-β

,β

-dimethyl-1,3-dioxane-2-ethanol diacrylate, 1,6-hexanediol propoxylate diacrylate, 3-hydroxy-2,2-dimethylpropyl 3-hydroxy-2,2-dimethylpropionate diacrylate, glycerol 1,3-diglycerolate diacrylate, glycerol dimethacrylate, mixture of isomers, tech., 85%, neopentyl glycol dimethacrylate, neopentyl glycol ethoxylate (1 EO/OH) diacrylate, trimethylolpropane benzoate diacrylate, 1,14-tetradecanediol dimethacrylate, tricyclo[5.2.1.0^2,6]decanedimethanol diacrylate, trimethylolpropane ethoxylate (1 EO/OH) methyl ether diacrylate, and trimethylolpropane triacrylate, tech.