Polymer-coated substrates for binding biomolecules and methods of making and using Thereof

• US 20110008912A1
• Filed: 09/21/2010
• Published: 01/13/2011
• Est. Priority Date: 11/24/2004
• Status: Abandoned Application

• Current Assignee: Anthony G. Frutos, Jinlin Peng, Joydeep Lahiri, Odessa N. Petzold
• Original Assignee: Anthony G. Frutos, Jinlin Peng, Joydeep Lahiri, Odessa N. Petzold
• Inventors: Peng, Jinlin, Lahiri, Joydeep, Frutos, Anthony G., Petzold, Odessa N.
• Application Number:
  US12/886,997
• Publication Number:
  US 20110008912A1
• Time in Patent Office: Days
• US Class Current: 436/501
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C03C 17/3405 with at least two coatings of organic materials C03C17/36, C03C17/42 take precedence
• CPC Class Codes: G01N 21/553 and using surface plasmons fluorescence excitation G01N21/648; enhanced Raman G01N21/658
• CPC Class Codes: G01N 21/648 using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/545 Synthetic resin
• CPC Class Codes: Y10T 428/26 Web or sheet containing structurally defined element or component, the element or component having a specified physical dimension
• CPC Class Codes: Y10T 428/269 including synthetic resin or polymer layer or component
• CPC Class Codes: Y10T 428/31504 Composite [nonstructural laminate]
• CPC Class Codes: Y10T 428/31536 Including interfacial reaction product of adjacent layers
• CPC Class Codes: Y10T 428/31663 As siloxane, silicone or silane
• CPC Class Codes: Y10T 428/31935 Ester, halide or nitrile of addition polymer

Abstract

Described herein are polymer-coated substrates for binding biomolecules and methods of making and using thereof.

Claims

1-55. -55. (canceled)

56. A method for preparing a label-independent surface modified microplate comprising:

attaching a first tie layer compound to the surface of the microplate;

attaching a first polymer to the first tie layer compound, the first polymer comprises a copolymer of maleic anhydride and a first monomer, the maleic anhydride in the first polymer comprises from about 5% to about 50 mol % of the first monomer; and

attaching a biomolecule to the first polymer, and the attaching is accomplished without a chemical activation step.

57. The method of claim 56, wherein the support comprises a plastic, a polymer or co-polymer substance, a ceramic, a glass, a metal, a crystalline material, a noble or semi-noble metal, a metallic or non-metallic oxide, a transition metal, or any combination thereof.

58. The method of claim 56, wherein the first tie layer is covalently attached to the outer surface of the support.

59. The method of claim 56, wherein the first tie layer is electrostatically attached to the outer surface of the support.

60. The method of claim 56, wherein the first tie-layer compound comprises one or more reactive functional groups.

61. The method of claim 60, wherein the functional group comprises an amino group, a thiol group, a hydroxyl group, a carboxyl group, an acrylic acid, an organic and inorganic acid, an ester, an anhydride, an aldehyde, an epoxide, their derivatives or salts thereof, or a combination thereof.

62. The method of claim 56, wherein the first tie layer compound comprises a straight or branched-chain aminosilane, aminoalkoxysilane, aminoalkylsilane, aminoarylsilane, aminoaryloxysilane, or a derivative or salt thereof.

63. The method of claim 56, wherein the first tie layer compound comprises 3-aminopropyl trimethoxysilane, N-(beta-aminoethyl)-3-aminopropyl trimethoxysilane, N-(beta-aminoethyl)-3-aminopropyl triethoxysilane, N′

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

64. The method of claim 56, wherein the first tie layer is derived from a polyamine.

65. The method of claim 64, wherein the first tie layer is derived from poly-lysine or polyethyleneimine.

66. The method of claim 56, wherein the first tie layer compound comprises a self-assembled monolayer.

67. The method of claim 56, wherein the first polymer is covalently attached to the tie layer.

68. The method of claim 56, wherein the first polymer is electrostatically attached to the tie layer.

70. The method of claim 56, wherein the first polymer comprises at least one electrophilic group susceptible to nucleophilic attack.

71. The method of claim 56, wherein the first polymer comprises at least one amine-reactive group.

72. The method of claim 71, wherein the amine-reactive group comprises an ester group, an epoxide group, or an aldehyde group.

73. The method of claim 71, wherein the amine-reactive group is an anhydride group.

76. The method of claim 56, wherein the amount of maleic anhydride is about 50% by stoichiometry of the first monomer.

77. The method of claim 56, wherein the first monomer comprises styrene, tetradecene, octadecene, methyl vinyl ether, triethylene glycol methyl vinyl ether, butylvinyl ether, divinylbenzene, ethylene, acrylamide, dimethylacrylamide, pyrolidone, a polymerizable oligo(ethylene glycol) or oligo(ethylene oxide), or a combination thereof.

78. The method of claim 56, wherein the first polymer comprises, poly(vinyl acetate-maleic anhydride), poly(styrene-co-maleic anhydride),poly(ethylene-alt-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(trithyleneglycol methyvinyl ether-co-maleic anhydride), or a combination thereof.

79. The method of claim 56, wherein the first polymer is poly(ethylene-alt-maleic anhydride).

80. The method of claim 56, wherein the first polymer comprises at least one monolayer.

81. The method of claim 56, wherein the first polymer has a thickness of about 10 Å

to about 2,000 Å

.

82. The method of claim 56, wherein after step (2), (3) attaching a second tie layer compound to the first polymer, and (4) attaching a second polymer to the second tie layer compound.

83. The method of claim 82, wherein the second tie layer compound is covalently and/or electrostatically attached to the first polymer, and the second polymer is covalently and/or electrostatically attached to the second tie layer compound.

84. The method of claim 82, wherein the second tie layer compound comprises a polyamine or polyol.

85. The method of claim 82, wherein the second tie layer compound comprises ethylene diamine, ethylene glycol, or an oligoethylene glycol diamine.

86. The method of claim 82, wherein the second tie layer compound comprises a diamine, a triamine, or a tetraamine.

87. The method of claim 82, wherein the second polymer comprises at least one anhydride group.

88. The method of claim 82, wherein the second polymer comprises polymaleic anhydride or a copolymer derived from maleic anhydride.

89. The method of claim 56, wherein after step (2), (3) attaching a linker to the first polymer.

90. The method of claim 89, wherein the linker comprises N-(5-amino-1-carboxypentyl)iminodiacetic acid.

91. The method of claim 56, wherein after step (2), (3) attaching a biomolecule to the first polymer.

92. The method of claim 91, wherein the biomolecule is attached to the first polymer by a chemical interaction, an electrostatic interaction, or a combination thereof.

93. The method of claim 91, wherein the biomolecule is covalently attached to the first polymer.

94. The method of claim 91, wherein the biomolecule comprises a natural or synthetic oligonucleotide, a natural or modified/blocked nucleotide/nucleoside, a nucleic acid (DNA) or (RNA), a peptide comprising natural or modified/blocked amino acid, an antibody, a hapten, a biological ligand, a protein membrane, a lipid membrane, a small molecule, or a cell.

95. The method of claim 91, wherein the biomolecule comprises a protein.

96. The method of claim 95, wherein the protein comprises a peptide, a fragment of a protein or peptide, a membrane-bound protein, or a nuclear protein.

97. The method of claim 91, wherein the biomolecule is attached to the substrate in a sufficient amount under about 1 hour.

98. The method of claim 91, wherein the biomolecule is attached to the substrate in a sufficient amount under about 0.5 hours.

99. The method of claim 91, wherein the biomolecule is attached to the first polymer at a pH of from about 0.5 to 1 pH units below the isoelectric point of the biomolecule.

100. The method of claim 56, wherein after step (2), (3) attaching a blocking agent to the first polymer.

101. The method of claim 100, wherein the blocking agent comprises a positively charged compound.

102. The method of claim 100, wherein the blocking agent comprises a positively charged dextran.

103. The method of claim 100, wherein the blocking agent is diethylaminethyl dextran.

104. The method of claim 91, wherein after step (3), (4) attaching a blocking agent to the first polymer.

105. The method of claim 56, wherein the first tie layer is aminopropylsilsesquioxane and the first polymer is poly(ethylene-alt-maleic anhydride).

106. The method of claim 56, wherein the substrate is a microplate or a slide.

107. An article produced by the method of claim 56.

69. (canceled)

74. (canceled)

75. (canceled)

108. A method for performing a label-independent assay of a ligand, comprising

contacting the ligand with a substrate comprising a first tie layer, a first polymer, the first polymer comprises a copolymer of maleic anhydride and a first monomer, the maleic anhydride in the first polymer comprises from about 5% to about 50 mol % of the first monomer and a biomolecule, wherein the tie layer attaches the first polymer to the substrate, the biomolecule is attached to the first polymer without a chemical activation step, and the ligand is bound to the biomolecules on the substrate after the contacting step;

and

detecting the bound ligand.

109. The method of claim 108, wherein the assay is a high-throughput assay.

110. The method of claim 108, wherein the ligand comprises a drug, an oligonucleotide, a nucleic acid, a protein, a peptide, an antibody, an antigen, a hapten, or a small molecule.

111. The method of claim 108, wherein the bound ligand is detected by fluorescence.

112. The method of claim 108, wherein the bound ligand is detected by surface plasmon resonance, a waveguide resonant grating system, or mass spectrometry.

113. The method of claim 108, wherein the substrate comprises a plastic, a polymer or co-polymer substance, a ceramic, a glass, a metal, a crystalline material, a noble or semi-noble metal, a metallic or non-metallic oxide, a transition metal, or any combination thereof.

114. The method of claim 108, wherein the first tie layer is covalently attached to the outer surface of the substrate.

115. The method of claim 108, wherein the first tie layer is electrostatically attached to the outer surface of the substrate.

116. The method of claim 108, wherein the first tie-layer compound comprises one or more reactive functional groups.

117. The method of claim 116, wherein the functional group comprises an amino group, a thiol group, a hydroxyl group, a carboxyl group, an acrylic acid, an organic and inorganic acid, an ester, an anhydride, an aldehyde, an epoxide, their derivatives or salts thereof, or a combination thereof.

118. The method of claim 108, wherein the first tie layer compound comprises a straight or branched-chain aminosilane, aminoalkoxysilane, aminoalkylsilane, aminoarylsilane, aminoaryloxysilane, or a derivative or salt thereof.

119. The method of claim 108, wherein the first tie layer compound comprises 3-aminopropyl trimethoxysilane, N-(beta-aminoethyl)-3-aminopropyl trimethoxysilane, N-(beta-aminoethyl)-3-aminopropyl triethoxysilane, N′

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

120. The method of claim 108, wherein the first tie layer is derived from a polyamine.

121. The method of claim 120, wherein the first tie layer is derived from poly-lysine or polyethyleneimine.

122. The method of claim 108, wherein the first tie layer compound comprises a self-assembled monolayer.

123. The method of claim 108, wherein the first polymer is covalently attached to the tie layer.

124. The method of claim 108, wherein the first polymer is electrostatically attached to the tie layer.

125. The method of claim 108, wherein the first polymer comprises a copolymer.

126. The method of claim 108, wherein the first polymer comprises at least one electrophilic group susceptible to nucleophilic attack.

127. The method of claim 108, wherein the first polymer comprises at least one amine-reactive group.

128. The method of claim 127, wherein the amine-reactive group comprises an ester group, an epoxide group, or an aldehyde group.

129. The method of claim 128, wherein the amine-reactive group is an anhydride group.

134. The method of claim 108, wherein the first polymer comprises, poly(vinyl acetate-maleic anhydride), poly(styrene-co-maleic anhydride), poly(ethylene-alt-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(trithyleneglycol methyvinyl ether-co-maleic anhydride), or a combination thereof.

135. The method of claim 108, wherein the first polymer is poly(ethylene-alt-maleic anhydride).

136. The method of claim 108, wherein the first polymer comprises at least one monolayer.

137. The method of claim 108, wherein the first polymer has a thickness of about 10 Å

to about 2,000 Å

.

138. The method of claim 108, wherein a second tie layer compound is attached to the first polymer, and a second polymer is attached to the second tie layer compound.

139. The method of claim 138, wherein a second tie layer compound is covalently and/or electrostatically attached to the first polymer, and a second polymer is covalently and/or electrostatically attached to the second tie layer compound.

140. The method of claim 138, wherein the second tie layer compound comprises a polyamine or polyol.

141. The method of claim 138, wherein the second tie layer compound comprises ethylene diamine, ethylene glycol, or an oligoethylene glycol diamine.

142. The method of claim 138, wherein the second tie layer compound comprises a diamine, a triamine, or a tetraamine.

143. The method of claim 138, wherein the second polymer comprises at least one anhydride group.

144. The method of claim 138, wherein the second polymer comprises polymaleic anhydride or a copolymer derived from maleic anhydride.

145. The method of claim 108, wherein the substrate further comprises a linker attached to the first polymer.

146. The method of claim 145, wherein the linker comprises N-(5-amino-1-carboxypentyl)iminodiacetic acid.

147. The method of claim 108, wherein the biomolecule is attached to the first polymer by a chemical interaction, an electrostatic interaction, or a combination thereof.

148. The method of claim 108, wherein the biomolecule is attached to the first polymer by a linker.

149. The method of claim 108, wherein the biomolecule is covalently attached to the first polymer.

150. The method of claim 108, wherein the biomolecule comprises a natural or synthetic oligonucleotide, a natural or modified/blocked nucleotide/nucleoside, a nucleic acid (DNA) or (RNA), a peptide comprising natural or modified/blocked amino acid, an antibody, a hapten, a biological ligand, a protein membrane, a lipid membrane, a small molecule, or a cell.

151. The method of claim 108, wherein the biomolecule comprises a protein.

152. The method of claim 151, wherein the protein comprises a peptide, a fragment of a protein or peptide, a membrane-bound protein, or a nuclear protein.

153. The method of claim 108, wherein the substrate further comprises a blocking agent attached to the first polymer.

154. The method of claim 153, wherein the blocking agent comprises a positively charged compound.

155. The method of claim 153, wherein the blocking agent comprises a positively charged dextran.

156. The method of claim 153, wherein the blocking agent is diethylaminethyl dextran.

157. The method of claim 108, wherein the first tie layer is aminopropylsilsesquioxane and the first polymer is poly(ethylene-alt-maleic anhydride).

158. The method of claim 108, wherein the substrate is a microplate or a slide.

160. The method of claim 108, wherein the bound ligand is detected by label independent detection.

161. The method of claim 108, wherein the substrate is a microplate, the first tie layer is derived from 3-aminopropyl trimethoxysilane, the first polymer is poly(ethylene-alt-maleic anhydride), and the bound ligand is detected by label independent detection.

130. (canceled)

132. The method of claim 130, wherein the amount of maleic anhydride is about 50% by stoichiometry of the first monomer.

133. The method of claim 130, wherein the first monomer comprises styrene, tetradecene, octadecene, methyl vinyl ether, triethylene glycol methyl vinyl ether, butylvinyl ether, divinylbenzene, ethylene, acrylamide, dimethylacrylamide, pyrolidone, a polymerizable oligo(ethylene glycol) or oligo(ethylene oxide), or a combination thereof.

131. (canceled)