Biosensor incorporating protein-immobilized membrane and method of immobilizing protein in biosensor
First Claim
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1. A biosensor, comprising:
- a substrate;
a capillary formed on the substrate for passage of a fluid to be analyzed;
an electrode formed on the substrate; and
a reagent portion formed on the substrate to cover the electrode;
wherein the reagent portion comprises;
an orientation-controlling layer formed on the electrode in contact therewith; and
a layer of protein that is immobilized on a side of the orientation-controlling layer opposite the electrode in direct contact with the orientation-controlling layer, the layer of protein containing a cytochrome or a cytochrome complex, and the cytochrome or cytochrome complex of the layer of protein being oriented toward the orientation-controlling layer; and
wherein the orientation-controlling layer contains a phospholipid polymer, the phospholipid polymer being 2-methacryloyloxyethyl phosphorylcholine polymer.
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Abstract
The present invention relates to a protein-immobilized membrane (14) including a cell membrane homologous layer (14A) and a protein (14B) immobilized to the cell membrane homologous layer (14A), where the protein contains cytochrome or a cytochrome complex. The present invention also relates to a method for forming a protein-immobilized membrane (14), and an enzyme-immobilized electrode and a biosensor (X1) provided with a protein-immobilized membrane (14). Preferably, the cell membrane homologous layer (14A) may contain a phospholipid polymer, and the protein (14B) may be CyGDH including an α subunit having a glucose dehydrogenase activity and cytochrome C having a function of electron transfer.
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Citations
21 Claims
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1. A biosensor, comprising:
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a substrate; a capillary formed on the substrate for passage of a fluid to be analyzed; an electrode formed on the substrate; and a reagent portion formed on the substrate to cover the electrode; wherein the reagent portion comprises; an orientation-controlling layer formed on the electrode in contact therewith; and a layer of protein that is immobilized on a side of the orientation-controlling layer opposite the electrode in direct contact with the orientation-controlling layer, the layer of protein containing a cytochrome or a cytochrome complex, and the cytochrome or cytochrome complex of the layer of protein being oriented toward the orientation-controlling layer; and wherein the orientation-controlling layer contains a phospholipid polymer, the phospholipid polymer being 2-methacryloyloxyethyl phosphorylcholine polymer. - View Dependent Claims (2, 3, 4)
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5. A biosensor, comprising:
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a substrate; a capillary formed on the substrate for passage of a fluid to be analyzed; an electrode formed on the substrate; and a reagent portion formed on the substrate to cover the electrode; wherein the reagent portion comprises; an orientation-controlling layer formed on the electrode in contact therewith; and a layer of protein that is immobilized on a side of the orientation-controlling layer opposite the electrode in direct contact with the orientation-controlling layer, the layer of protein containing a cytochrome or a cytochrome complex, and the cytochrome or cytochrome complex of the layer of protein being oriented toward the orientation-controlling layer; and wherein the orientation-controlling layer contains a silane coupling agent for bonding the orientation-controlling layer to the electrode. - View Dependent Claims (6, 7, 8)
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9. A method for immobilizing a protein in a biosensor, the biosensor comprising:
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a substrate; a capillary formed on the substrate for passage of a fluid to be analyzed; and
an electrode formed on the substrate;the method comprising; forming an orientation-controlling layer on a portion of the electrode in contact therewith; and causing a layer of protein to immobilize on a side of the orientation-controlling layer opposite the electrode in direct contact with the orientation-controlling layer, the layer of protein containing cytochrome or a cytochrome complex, the cytochrome or cytochrome complex of the layer of protein being oriented toward the orientation-controlling layer; and wherein the orientation-controlling layer contains a phospholipid polymer, the phospholipid polymer is 2-methacryloyloxyethyl phosphorylcholine polymer. - View Dependent Claims (10, 11, 12, 13, 14)
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15. A method for immobilizing a protein in a biosensor, the biosensor comprising:
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a substrate; a capillary formed on the substrate for passage of a fluid to be analyzed; and
an electrode formed on the substrate;the method comprising; forming an orientation-controlling layer on a portion of the electrode in contact therewith; and causing a layer of protein to immobilize on a side of the orientation-controlling layer opposite the electrode in direct contact with the orientation-controlling layer, the layer of protein containing cytochrome or a cytochrome complex, the cytochrome or cytochrome complex of the layer of protein being oriented toward the orientation-controlling layer; and wherein the orientation-controlling layer is formed to contain a silane coupling agent for bonding the orientation-controlling layer to the electrode. - View Dependent Claims (16, 17, 18, 19, 20, 21)
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Specification