SUPPORTS FOR ASSAYING ANALYTES AND METHODS OF MAKING AND USING THEREOF

• US 20090137425A1
• Filed: 01/22/2009
• Published: 05/28/2009
• Est. Priority Date: 12/29/2005
• Status: Active Grant

• Current Assignee: Anthony G. Frutos, David Henry
• Original Assignee: Anthony G. Frutos, David Henry
• Inventors: Frutos, Anthony G., Henry, David
• Granted Patent: US 7,981,665 B2
• Time in Patent Office: Days
• US Class Current: 506/14
• CPC Class Codes: B01J 2219/00527 Sheets
• 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 2219/0063 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 2219/0072 Organic compounds
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/00731 Saccharides
• CPC Class Codes: B01J 2219/00736 Non-biologic macromolecules, e.g. polymeric compounds
• CPC Class Codes: B01J 2219/0074 Biological products
• CPC Class Codes: C08F 210/02 Ethene
• CPC Class Codes: C08F 222/06 Maleic anhydride
• CPC Class Codes: C08F 8/00 Chemical modification by after-treatment graft polymers, block polymers, crosslinking with unsaturated monomers or with polymers C08F251/00 - C08F299/00; of conjugated diene rubbers C08C
• CPC Class Codes: C08F 8/32 by reaction with amines
• CPC Class Codes: G01N 33/54353 : with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: Y10T 436/143333 : Saccharide [e.g., DNA, etc.]

Abstract

Described herein are supports for assaying an analyte and methods of making and using thereof.

Claims

1. A support for performing an assay, comprising a substrate and a binding polymer directly or indirectly attached to the substrate, wherein the binding polymer has a plurality of reactive groups capable of attaching to a biomolecule and a plurality of ionizable groups, wherein the ratio of reactive groups to ionizable groups is from 0.5 to 10, wherein the binding polymer does not contain a photoreactive group.

2. The support of claim 1, wherein the substrate comprises a plastic, a polymeric or co-polymeric substance, a ceramic, a glass, a metal, a crystalline material, a noble or semi-noble metal, a metallic or non-metallic oxide, an inorganic oxide, an inorganic nitride, a transition metal, or any combination thereof.

3. The support of claim 1, wherein the substrate comprises gold.

4. The support of claim 1, wherein the substrate comprises glass with a layer comprising Ta₂O₅, Nb₂O₅, TiO₂, Al₂O₃, SiO₂, silicon nitride or a mixture thereof, wherein the layer is adjacent to the surface of the glass.

5. The support of claim 1, wherein the substrate is a microplate or a slide.

6. The support of claim 1, wherein the binding polymer is directly attached to the substrate.

7. The support of claim 1, wherein the substrate is amine-modified.

8. The support of claim 1, wherein the substrate is modified with an aminosilane or a polymer comprising at least one amino group.

9. The support of claim 1, wherein the substrate is modified with poly-lysine, polyethyleneimine, poly(allyl)amine, or silylated polyethyleneimine.

10. The support of claim 1, wherein the binding polymer is indirectly attached to the substrate by a tie layer.

11. The support of claim 10, wherein the tie layer is covalently attached to the outer surface of the substrate.

12. The support of claim 10, wherein the tie layer is electrostatically attached to the outer surface of the substrate

13. The support of claim 10, wherein the tie layer is derived from a compound comprising a straight or branched-chain aminosilane, aminoalkoxysilane, aminoalkylsilane, aminoarylsilane, aminoaryloxysilane, or a derivative or salt thereof.

14. The support of claim 10, wherein the tie layer is derived from a compound comprising 3-aminopropyl trimethoxysilane, N-(beta-aminoethyl)-3-aminopropyl trimethoxysilane, N-(beta-aminoethyl)-3-aminopropyl triethoxysilane, N′

-(beta-aminoethyl)₃-aminopropyl methoxysilane, or aminopropylsilsesquixoane.

15. The support of claim 1, wherein the substrate is modified with an aminothiol.

16. The support of claim 15, wherein the aminothiol comprises 1-amino-1-undecanethiol hydrochloride.

17. The support of claim 10, wherein the binding polymer is covalently attached to the tie layer.

18. The support of claim 10, wherein the binding polymer is electrostatically attached to the tie layer.

19. The support of claim 1, wherein the reactive group comprises an anhydride group, an epoxy group, an aldehyde group, an activated ester, an isocyanate, an isothiocyanate, a sulfonyl chloride, a carbonate, an aryl or alkyl halide, an aziridine, or a maleimide.

20. The support of claim 1, wherein the ionizable group can be converted to a positively charged group.

21. The support of claim 1, wherein the ionizable group can be converted to a negatively charged group.

22. The support of claim 20, wherein the positively charged group comprises an ammonium group.

23. The support of claim 21, wherein the negatively charged group comprises a carboxylate, sulfonate, or phosphonate group.

24. The support of claim 1, wherein the binding polymer comprises a copolymer.

25. The support of claim 24, wherein the copolymer is derived from maleic anhydride and a first monomer.

26. The support of claim 25, wherein the first monomer comprises styrene, tetradecene, octadecene, methyl vinyl ether, triethylene glycol methyl vinyl ether, butylvinyl ether, divinylbenzene, ethylene, acrylamide, vinyl pyrolidone, dimethylacrylamide, a polymerizable oligo(ethylene glycol) or oligo(ethylene oxide), propylene, isobutylene, vinyl acetate, methacrylate, acrylate, methacrylamide, or a combination thereof.

27. The support of claim 1, wherein the binding polymer comprises poly(vinyl acetate-maleic anhydride), poly(styrene-co-maleic anhydride), poly(isobutylene-alt-maleic anhydride), poly(maleic anhydride-alt-1-octadecene), poly(maleic anhydride-alt-1-tetradecene), poly(maleic anhydride-alt-methyl vinyl ether), poly(triethyleneglycol methylvinyl ether-co-maleic anhydride), or any combination thereof.

28. The support of claim 1, wherein the binding polymer comprises poly(ethylene-alt-maleic anhydride).

29. The support of claim 1, wherein one or more biomolecules are attached to the reactive groups of the binding polymer.

30. The support of claim 29, wherein the biomolecule comprises a natural, synthetic or modified oligonucleotide, a natural or modified nucleotide or nucleoside, a nucleic acid (DNA) or (RNA) or fragment thereof, a peptide comprising natural or modified amino acid, an antibody, a hapten, a biological ligand, a chelate, an aptamer, a lipid, a saccharide, a small molecule, a lectin, a modified polysaccharide, a synthetic composite macromolecule, a functionalized nanostructure, a synthetic polymer, a fluorophore, a chromophore, or a cell.

31. The support of claim 29, wherein the biomolecule comprises an oligonucleotide.

32. The support of claim 29, wherein the biomolecule comprises DNA or a fragment thereof.

33. The support of claim 29, wherein the biomolecule comprises RNA or a fragment thereof.

34. The support of claim 29, wherein the biomolecule comprises a protein, a peptide, or fragments thereof.

35. The support of claim 29, wherein the biomolecule is covalently attached to the binding polymer.

36. The support of claim 29, wherein the biomolecule is electrostatically attached to the binding polymer.

37. The support of claim 1, wherein a plurality of biomolecules are present on the support and wherein the biomolecules are on discrete and defined locations on the support to produce an array.

38. The support of claim 37, wherein the array comprises at least 96 distinct and defined locations.

39. The support of claim 37, wherein the support comprises at least 384 distinct and defined locations.

40. The support of claim 1, wherein the ratio of reactive groups to ionizable groups is from 0.5 to 5.0.

41. The support of claim 1, wherein the ratio of reactive groups to ionizable groups is from 0.67 to 3.0.

42. The support of claim 1, wherein the substrate comprises a gold chip, the binding polymer comprises poly(ethylene-alt-maleic anhydride) indirectly attached to the substrate by an aminothiol, and the ratio of reactive groups to ionizable groups in the binding polymer is from 0.67 to 3.0.

43. The support of claim 1, wherein the substrate comprises a glass substrate with a layer comprising Ta₂O₅, Nb₂O₅, TiO₂, Al₂O₃, silicon nitride, SiO₂ or a mixture thereof, the binding polymer comprises poly(ethylene-alt-maleic anhydride) indirectly attached to the substrate by a tie layer, wherein the tie layer is derived from aminopropylsilane, and the ratio of reactive groups to ionizable groups in the binding polymer is from 0.67 to 3.0.

44. A method for performing an assay of a sample comprising an analyte, the method comprising:

a. contacting a support with a sample comprising the analyte, wherein the support comprises a substrate and a binding polymer directly or indirectly attached to the substrate, wherein the binding polymer has a plurality of reactive groups capable of attaching to a biomolecule and a plurality of ionizable groups, wherein the ratio of reactive groups to ionizable groups is from 0.5 to 10, wherein the binding polymer does not contain a photoreactive group, and

b. detecting the bound analyte.

45. The method of claim 44, wherein the substrate comprises a plastic, a polymeric or co-polymeric substance, a ceramic, a glass, a metal, a crystalline material, a noble or semi-noble metal, a metallic or non-metallic oxide, an inorganic oxide, an inorganic nitride, a transition metal, or any combination thereof.

46. The method of claim 44, wherein the substrate comprises gold.

47. The method of claim 44, wherein the substrate comprises SiO₂ with a layer comprising Ta₂O₅, Nb₂O₅, TiO₂, Al₂O₃, silicon nitride or a mixture thereof, wherein the layer is adjacent to the surface of the SiO₂.

48. The method of claim 44, wherein the substrate is a microplate or a slide.

49. The method of claim 44, wherein the binding polymer is directly attached to the substrate.

50. The method of claim 44, wherein the substrate is amine-modified.

51. The method of claim 44, wherein the substrate is modified with an aminosilane or a polymer comprising at least one amino group.

52. The method of claim 44, wherein the substrate is modified with polysine, polyethyleneimine, poly(allyl)amine, or silylated polyethyleneimine.

53. The method of claim 44, wherein the binding polymer is indirectly attached to the substrate by a tie layer.

54. The method of claim 53, wherein the tie layer is covalently attached to the outer surface of the substrate.

55. The method of claim 53, wherein the tie layer is electrostatically attached to the outer surface of the substrate

56. The method of claim 53, wherein the tie layer is derived from a compound comprising a straight or branched-chain aminosilane, aminoalkoxysilane, aminoalkylsilane, aminoarylsilane, aminoaryloxysilane, or a derivative or salt thereof.

57. The method of claim 53, wherein the lie layer is derived from a compound comprising 3-aminopropyl trimethoxysilane, N-(beta-aminoethyl)-3-aminopropyl trimethoxysilane, N-(beta-aminoethyl)-3-aminopropyl triethoxysilane, N′

-(beta-aminoethyl)-3-aminopropyl methoxysilane, or aminopropylsilsesquixoane.

58. The method of claim 44, wherein the substrate is modified with an aminothiol.

59. The method of claim 58, wherein the aminothiol comprises 11-amino-1-undecanethiol hydrochloride.

60. The method of claim 53, wherein the binding polymer is covalently attached to the tie layer.

61. The method of claim 53, wherein the binding polymer is electrostatically attached to the tie layer.

62. The method of claim 44, wherein the reactive group comprises an anhydride group, an epoxy group, an aldehyde group, an activated ester, an isocyanate, an isothiocyanate, a sulfonyl chloride, a carbonate, an aryl or alkyl halide, an aziridine, or a maleimide.

63. The method of claim 44, wherein the ionizable group can be converted to a positively charged group.

64. The method of claim 44, wherein the ionizable group can be converted to a negatively charged group.

65. The method of claim 63, wherein the positively charged group comprises an ammonium group.

66. The method of claim 64, wherein the negatively charged group comprises a carboxylate, sulfonate, or phosphonate group.

67. The method of claim 44, wherein the binding polymer comprises a copolymer.

70. The method of claim 44, wherein the binding polymer comprises poly(vinyl acetate-maleic anhydride), poly(styrene-co-maleic anhydride), poly(isobutylene-alt-maleic anhydride), poly(maleic anhydride-alt-1-octadecene), poly(maleic anhydride-alt-1-tetradecene), poly(maleic anhydride-alt-methyl vinyl ether), poly(triethyleneglycol methylvinyl ether-co-maleic anhydride), or any combination thereof.

71. The method of claim 44, wherein the binding polymer comprises poly(ethylene-alt-maleic anhydride).

72. The method of claim 44, wherein one or more biomolecules are attached to the reactive groups of the binding polymer.

73. The method of claim 72, wherein the biomolecule comprises a natural synthetic or modified oligonucleotide, a natural or modified nucleotide or nucleoside, a nucleic acid (DNA) or (RNA) or fragment thereof, a peptide comprising natural or modified amino acid, an antibody, a hapten, a biological ligand, a chelate, an aptamer, a lipid, a saccharide, a small molecule, a lectin, a modified polysaccharide, a synthetic composite macromolecule, a functionalized nanostructure, a synthetic polymer, a fluorophore, a chromophore, or a cell.

74. The method of claim 72, wherein the biomolecule comprises an oligonucleotide.

75. The method of claim 72, wherein the biomolecule comprises DNA or a fragment thereof.

76. The method of claim 72, wherein the biomolecule comprises RNA or a fragment thereof.

77. The method of claim 72, wherein the biomolecule comprises a protein, a peptide, or fragments thereof.

78. The method of claim 72, wherein the biomolecule is covalently attached to the binding polymer.

79. The method of claim 72, wherein the biomolecule is electrostatically attached to the binding polymer.

80. The method of claim 44, wherein a plurality of biomolecules are present on the support and wherein the biomolecules are on discrete and defined locations on the support to produce an array.

81. The method of claim 80, wherein the array comprises at least 96 distinct and defined locations.

82. The method of claim 80, wherein the support comprises at least 384 distinct and defined locations.

83. The method of claim 44, wherein the ratio of reactive groups to ionizable groups is from 0.5 to 5.0.

84. The method of claim 44, wherein the ratio of reactive groups to ionizable groups is from 0.67 to 3.0.

85. The method of claim 44, wherein the substrate comprises a gold chip, the binding polymer comprises poly(ethylene-alt-maleic anhydride) indirectly attached to the substrate by an aminothiol, and the ratio of reactive groups to ionizable groups in the binding polymer is from 0.67 to 3.0.

86. The method of claim 44, wherein the substrate comprises a glass substrate with a layer comprising Ta₂O₅, Nb₂O₅, TiO₂, Al₂O₃, silicon nitride, SiO2 or a mixture thereof, the binding polymer comprises poly(ethylene-alt-maleic anhydride) indirectly attached to the substrate by a tie layer, wherein the tie layer is derived from aminopropylsilane, and the ratio of reactive groups to ionizable groups in the binding polymer is from 0.67 to 3.0.

87. The method of claim 44, wherein the assay is a high-throughput assay.

88. The method of claim 44, wherein the analyte comprises a drug, an oligonucleotide, a nucleic acid, a protein, a peptide, an antibody, an antigen, a hapten, saccharide, lipid, a small molecule, or a mixture thereof.

89. The method of claim 44, wherein the bound analyte is detected by fluorescence.

90. The method of claim 44, wherein the bound analyte is detected by a refractive index sensor, an acoustic wave sensor, or a mass sensor such as mass spectrometry or a quartz crystal microbalance.

68. The method of claim 68, wherein the copolymer is derived from maleic anhydride and a first monomer.

69. The method of claim 68, wherein the first monomer comprises styrene, tetradecene, octadecene, methyl vinyl ether, triethylene glycol methyl vinyl ether, butylvinyl ether, divinylbenzene, ethylene, acrylamide, vinyl pyrolidone, dimethylacrylamide, a polymerizable oligo(ethylene glycol) or oligo(ethylene oxide), propylene, isobutylene, vinyl acetate, methacrylate, acrylate, methacrylamide, or a combination thereof.

91. A method for making a support for performing an assay, comprising attaching a binding polymer directly or indirectly to a substrate, the binding polymer has a plurality of reactive groups capable of attaching to a biomolecule and a plurality of ionizable groups, the ratio of reactive groups to ionizable groups is from 0.5 to 10.0, and the binding polymer does not contain a photoreactive group.

92. The method of claim 91, wherein prior to attaching the binding polymer to the substrate, inactivating one or more reactive groups with a blocking agent.

93. The method of claim 92, wherein the blocking agent comprises a nucleophilic group.

94. The method of claim 92, wherein the blocking agent comprises an alkyl amine, an alkylhydroxy amine, an alkoxyalkyl amine, an alcohol, an alkyl thiol, water, or H₂S.

95. The method of claim 92, wherein the blocking agent comprises water, ammonia, 2-(2-aminoethoxy)ethanol, N,N-dimethyl ethylenediamine, ethanolamine, ethylenediamine, hydroxyl amine, methoxyethyl amine, ethyl amine, isopropyl amine, butyl amine, propyl amine, hexyl amine, 2-amino-2-methyl-1-propanol, 2-(2-aminoethyl amino) ethanol, 2-(2-aminoethoxy)ethanol, dimethylethanolamine, dibutyl ethanolamine, 1-amino-2-propanol, polyethylene glycol, polypropylene glycol, 4,7,10-trioxa-1,13-tridecanediamine, polyethylene glycol or an amine-terminated-polyethylene glycol, Trizma hydrochloride, or any combination thereof.

96. The method of claim 92, wherein the blocking agent comprises methoxyethyl amine.

97. The method of claim 91, wherein the substrate comprises a plastic, a polymeric or co-polymeric substance, a ceramic, a glass, a metal, a crystalline material, a noble or semi-noble metal, a metallic or non-metallic oxide, an inorganic oxide, an inorganic nitride, a transition metal, or any combination thereof.

98. The method of claim 91, wherein the substrate comprises gold.

99. The method of claim 91, wherein the substrate comprises SiO₂ with a layer comprising Ta₂O₅, Nb₂O₅, TiO₂, Al₂O₃, silicon nitride or a mixture thereof, wherein the layer is adjacent to the surface of the SiO₂.

100. The method of claim 91, wherein the binding polymer comprises poly(vinyl acetate-maleic anhydride), poly(styrene-co-maleic anhydride), poly(isobutylene-alt-maleic anhydride), poly(maleic anhydride-alt-1-octadecene), poly(maleic anhydride-alt-1-tetradecene), poly(maleic anhydride-alt-methyl vinyl ether), poly(triethyleneglycol methylvinyl ether-co-maleic anhydride), or any combination thereof.

101. The method of claim 91, wherein the binding polymer comprises poly(ethylene-all-maleic anhydride).

102. The method of claim 91, wherein the ratio of reactive groups to ionizable groups is from 0.5 to 5.0.

103. The method of claim 91, wherein the ratio of reactive groups to ionizable groups is from 0.67 to 3.0.

104. The method of claim 91, wherein the substrate comprises a gold chip, the binding polymer comprises poly(ethylene-alt-maleic anhydride) indirectly attached to the substrate by an aminothiol, and the ratio of reactive groups to ionizable groups in the binding polymer is from 0.67 to 3.0.

105. The method of claim 104, wherein prior to attachment to the substrate, the binding polymer is blocked with methoxyethyl amine.

106. The method of claim 91, wherein the substrate comprises a glass substrate with a layer comprising Ta₂O₅, Nb₂O₅, TiO₂, Al₂O₃, silicon nitride, SiO₂ or a mixture thereof, the binding polymer comprises poly(ethylene-alt-maleic anhydride) indirectly attached to the substrate by a tie layer, wherein the tie layer is derived from aminopropylsilane, and the ratio of reactive groups to ionizable groups in the binding polymer is from 0.67 to 3.0.

107. The method of claim 106, wherein prior to attachment to the substrate, the binding polymer is blocked with methoxyethyl amine.

109. A support made by the method of claim 91.

111. The support of claim 109 wherein the substrate comprises a cell culture surface.

112. The support of claim 109 wherein the assay comprises a cell based assay.

113. The support of claim 109 wherein the biomolecule comprises a cell.

108. A method for making a support for performing an assay, comprising

a. attaching a binding polymer directly or indirectly to a substrate, the binding polymer has a plurality of reactive groups capable of attaching to a biomolecule, the binding polymer does not contain a photoreactive group, and

b. converting the reactive groups to ionizable groups so that the ratio of reactive groups to ionizable groups is from 0.5 to 10.0.

110. A support made by the method of claim 108.