Photocrosslinked hydrogel surface coatings

• US 20070082019A1
• Filed: 02/20/2004
• Published: 04/12/2007
• Est. Priority Date: 02/21/2003
• Status: Abandoned Application

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Ciphergen Biosystems Incorporated
• Inventors: Boschetti, Egisto, Agroskin, Yury, Huang, Wenxi
• Application Number:
  US10/546,173
• Publication Number:
  US 20070082019A1
• Time in Patent Office: Days
• US Class Current: 424/423
• CPC Class Codes: A61L 27/52 Hydrogels or hydrocolloids
• CPC Class Codes: H01J 49/0418 for laser desorption, e.g. matrix-assisted laser desorption/ionisation [MALDI] plates or surface enhanced laser desorption/ionisation [SELDI] plates

Abstract

A hydrogel layer is applied to a substrate advantageously when the layer is formed in situ, using a polymeric or copolymeric precursor that includes, respectively, monomer subunits that have a photocrosslinkable functionality and monomer subunits that have a chemically selective functionality for binding a biomolecular analyte, such as a protein. A hydrogel-coated substrate thus obtained is particularly useful as a probe for mass spectroscopic analysis, including SELDI analysis. Hydrogel particles also can be used for SELDI analysis.

Claims

1. A copolymeric hydrogel precursor comprising:

(a) first monomeric subunits that comprise a photocrosslinkable functionality and

(b) second monomeric subunits that comprise a chemically selective functionality for binding a protein,

wherein the amounts of first and second monomeric subunits provide the copolymeric hydrogel precursor with the ability to be photocrosslinked into a hydrogel and the ability for the hydrogel to be selectively reactive with protein under aqueous conditions, whereby protein becomes bound to the chemically selective functionality.

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22. A polymeric hydrogel precursor comprising photocrosslinkable functionality and chemically selective functionality, wherein the precursor is prepared by functionalizing a prefunctionalized polymeric hydrogel precursor with photocrosslinkable functionality and with chemically selective functionality, wherein the amounts of photocrosslinkable functionality and chemically selective functionality provide the hydrogel precursor with the ability to be photocrosslinked into the hydrogel and the ability for the hydrogel to be selectively reactive with protein under aqueous conditions, whereby protein becomes bound to the chemically selective functionality.

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25. A copolymeric hydrogel precursor prepared by copolymerization of monomeric subunits comprising:

(a) first monomeric subunits that comprise a first free radical copolymerization functionality and a photocrosslinkable functionality and

(b) second monomeric subunits that comprise a second free radical copolymerization functionality and a chemically selective functionality for binding a protein,

wherein the amounts of first and second monomeric subunits provide, upon copolymerization, the polymeric hydrogel precursor with the ability to be photocrosslinked into the hydrogel and the ability to be selectively reactive with protein under aqueous conditions, whereby protein becomes bound to the chemically selective functionality.

26. A photocrosslinkable hydrogel precursor composition for selective interaction with protein under aqueous conditions consisting essentially of at least one hydrogel precursor polymer consisting essentially of (i) first comonomeric subunits that comprise a photocrosslinkable functionality and (ii) second comonomeric subunits that comprise a chemically selective functionality for interaction with a biomolecular analyte, wherein the photocrosslinkable hydrogel precursor composition is substantially free of photoinitiator.

27. A method for functionalizing a surface with copolymeric hydrogel, comprising:

(A) providing (i) a substrate presenting a surface and (ii) a copolymeric hydrogel precursor that comprises (a) first comonomeric subunits that comprise a photocrosslinkable functionality and (b) second comonomeric subunits that comprise a chemically selective functionality for binding a biomolecular analyte;

(B) contacting the copolymeric hydrogel precursor and the surface to form a layer of the copolymeric hydrogel precursor on the surface; and

(C) photocrosslinking at least some of the copolymeric hydrogel precursor layer to form hydrogel in contact with the surface.

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36. A substrate that comprises a substrate surface and a hydrogel thereon, wherein the hydrogel comprises (i) a photocrosslinked functionality and (ii) a chemically selective functionality for binding a biomolecular analyte, wherein the hydrogel is substantially free of photoinitiator, and wherein the amount of the chemically selective functionality is sufficient for binding the biomolecular analyte.

37. The substrate according to claim 36, wherein the substrate surface is the surface of a primer layer.

38. The substrate according to claim 36, wherein the substrate surface is planar.

39. The substrate according to claim 38, wherein the substrate surface is the surface of a primer layer and wherein the hydrogel is a uniform layer.

40. The substrate according to claim 38, wherein the substrate surface is the surface of a primer layer, and the hydrogel is in the form of discreet spots.

41. The substrate according to claim 36, wherein the hydrogel is covalently bound to the substrate surface.

42. The substrate according to claim 41, wherein the hydrogel comprises a photocrosslinked benzophenone functionality.

43. The substrate according to claim 36, wherein the hydrogel is a uniform layer having a layer thickness of 10 nm to 10 microns.

44. The substrate according to claim 36, wherein the hydrogel is a uniform layer having a layer thickness of about 2 microns or less.

45. The substrate according to claim 36, wherein the hydrogel is in the form of discreet spots and has a spot thickness of 10 nm to 10 microns.

46. The substrate according to claim 36, wherein said substrate is a biochip and said hydrogel is covalently bound to the surface.

47. The substrate according to claim 46, wherein the substrate comprises a supporting layer that comprises a material selected from the group consisting of aluminum, silicon, glass, metal oxide, metal, polymer and composite, and wherein said surface is a surface of a primer layer that is supported on the supporting layer.

48. The substrate according to claim 46, wherein the substrate comprises a supporting layer that comprises polymer or composite.

49. The substrate according to claim 46, wherein the substrate comprises a supporting layer that comprises polymer and is electrically conductive.

50. The substrate according to claim 46, wherein the primer layer is a hydrophobic primer layer.

51. The substrate according to claim 50, wherein the primer layer is a silane primer layer, a hydrocarbon silane primer layer, a fluorinated silane primer layer, a mixed fluorinated/hydrocarbon silane primer layer, or a polymeric primer layer.

52. The substrate according to claim 37, wherein the primer layer is about 4 angstroms to about 3 microns thick.

53. The substrate according to claim 37, wherein the primer layer is about 4 angstroms to about 10 nm thick.

54. The substrate according to claim 47, wherein the hydrogel is present on the surface only in one or more discreet spots.

55. The substrate according to claim 54, wherein the hydrogel is present as a plurality of discreet spots, each having at least one lateral dimension that is about 100 nm to about 3 mm.

56. The substrate according to claim 55, wherein said lateral dimension is about 500 nm to about 500 microns.

57. The substrate according to claim 46, wherein the hydrogel is a uniform layer on the substrate surface having an average layer thickness of 5 nm to 10 microns.

58. The substrate according to claim 46, wherein the hydrogel is a copolymeric hydrogel, a dextran derivative, or a derivative of poly(2-hydroxyethyl methacrylate) or copolymer thereof.

59. The substrate according to claim 46, wherein the hydrogel is comprised of photocrosslinked benzophenone, diazo ester, aryl azide, or diazirine functionality.

60. The substrate according to claim 46, wherein said chemically selective functionality is a nucleophilic or electrophilic group.

61. The substrate according to claim 46, wherein said chemically selective functionality is an anionic or cationic group.

62. The substrate according to claim 46, wherein said chemically selective functionality is a carboxylic acid, amino, or quaternary amino group.

63. The substrate according to claim 46, wherein said hydrogel is a water-swellable polymer that comprises a linear, carbon backbone that has been crosslinked.

64. The substrate according to claim 46, wherein the hydrogel is a copolymer prepared by crosslinking of a precursor copolymer comprised of carboxylic acid-containing side groups and benzophenone-containing side groups.

65. The substrate according to claim 36, wherein the hydrogel is free of photoinitiator.

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75. A copolymeric hydrogel precursor comprising:

(a) first monomeric subunits that comprise a photocrosslinkable functionality and

(b) second monomeric subunits that comprise a chemically selective functionality for binding a biopolymer,

wherein the amounts of first and second monomeric subunits provide the copolymeric hydrogel precursor with the ability to be photocrosslinked into the hydrogel and the ability for the hydrogel to be selectively reactive with biopolymer under aqueous conditions, whereby biopolymer becomes bound to the chemically selective functionality.

76. A copolymeric hydrogel precursor comprising:

(a) first monomeric subunits that comprise a photocrosslinkable functionality and

(b) second monomeric subunits that comprise a chemically selective functionality for interacting with a biopolymer,

wherein the amounts of first and second monomeric subunits provide the copolymeric hydrogel precursor with the ability to be photocrosslinked into the hydrogel and the ability for the hydrogel to be selectively interactive with biopolymer under aqueous conditions, whereby biopolymer becomes adsorbed to the chemically selective functionality.

77. A copolymeric hydrogel precursor comprising:

(a) first monomeric subunits that comprise a photocrosslinkable functionality, and

(b) third monomeric subunits that comprise an energy absorbing moiety.

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