Measuring chip for surface plasmon resonance biosensor and method for producing the same

Abstract

An objective of the present invention is to provide a measuring chip for a surface plasmon resonance sensor that can detect a small amount of target substances in high sensitivity. The present invention provides a measuring chip for a surface plasmon resonance sensor comprising a metal layer, one or more plasma polymerization layers formed on said metal layer, and a biologically active substance immobilized on the surface of said plasma polymerization layer.

1 . A measuring chip for a surface plasmon resonance sensor comprising a metal layer and one or more plasma polymerization layers formed on said metal layer.  
     
     
         2 . A measuring chip for a surface plasmon resonance sensor comprising a metal layer, one or more plasma polymerization layers formed on said metal layer, and a nucleic acid immobilized on the surface of said plasma polymerization layer.  
     
     
         3 . A measuring chip for a surface plasmon resonance sensor according to  claim 2 , wherein said nucleic acid is DNA, RNA or PNA.  
     
     
         4 . A measuring chip for a surface plasmon resonance sensor comprising a metal layer, one or more plasma polymerization layers formed on said metal layer, and a non-immune protein immobilized on the surface of said plasma polymerization layer.  
     
     
         5 . A measuring chip for a surface plasmon resonance sensor according to  claim 4 , wherein said non-immune protein is either avidin, streptoavidin, biotin or a receptor.  
     
     
         6 . A measuring chip for a surface plasmon resonance sensor comprising a metal layer, one or more plasma polymerization layers formed on said metal layer, and an immunoglobulin-binding protein immobilized on the surface of said plasma polymerization layer.  
     
     
         7 . A measuring chip for a surface plasmon resonance sensor according to  claim 6 , wherein said immunoglobulin-binding protein is protein A, protein G, or a rheumatoid factor.  
     
     
         8 . A measuring chip for a surface plasmon resonance sensor comprising a metal layer, one or more plasma polymerization layers formed on said metal layer, and a sugar-binding protein immobilized on the surface of said plasma polymerization layer.  
     
     
         9 . A measuring chip for a surface plasmon resonance sensor according to  claim 8 , wherein said sugar-binding protein is lectin.  
     
     
         10 . A measuring chip for a surface plasmon resonance sensor comprising a metal layer, one or more plasma polymerization layers formed on said metal layer, and a sugar-recognizing sugar chain immobilized on the surface of said plasma polymerization layer.  
     
     
         11 . A measuring chip for a surface plasmon resonance sensor comprising a metal layer, one or more plasma polymerization layers formed on said metal layer, and a fatty acid or a fatty acid ester immobilized on the surface of said plasma polymerization layer.  
     
     
         12 . A measuring chip for a surface plasmon resonance sensor according to  claim 11 , wherein said fatty acid or fatty acid ester is either stearic acid, alachidic acid, behenic acid, ethyl stearate, ethyl arachidate, or ethyl behanate.  
     
     
         13 . A measuring chip for a surface plasmon resonance sensor comprising a metal layer, one or more plasma polymerization layers formed on said metal layer, and a polypeptide or oligopeptide having ligand-binding activity immobilized on the surface of said plasma polymerization layer.  
     
     
         14 . A measuring chip for a surface plasmon resonance sensor according to  claim 13 , wherein said polypeptide or oligopeptide is produced by using genetic engineering techniques or chemical synthesis methods.  
     
     
         15 . A measuring chip according to any one of  claims 1  to  14 , which further comprises an optically transparent substrate on which said metal layer is formed.  
     
     
         16 . A measuring chip according to any one of  claims 1  to  15 , wherein said plasma polymerization layer comprises a compound having one or more groups selected from the group consisting of —COOH, —CHO, —SH, —NH 2 , —OH, ═NH, —CONH 2 , —NCO, —CH═CH 2 , ═C═O, and  
       
         
           
           
               
               
           
         
       
     
     
         17 . A measuring chip according to any one of  claims 1  to  15 , wherein said plasma polymerization layer consists of two or more layers.  
     
     
         18 . A measuring chip according to any one of  claims 1  to  15 , wherein said monomer material of a plasma polymerization layer is a compound containing nitrogen.  
     
     
         19 . A measuring chip according to  claim 18 , wherein said compound containing nitrogen is CH 3 —(CH 2 ) n —NH 2  (wherein n is an integer from 1 to 6) and/or NH 2 —(CH 2 ) n —NH 2  (wherein n is an integer from 1 to 6).  
     
     
         20 . A measuring chip according to  claim 18 , wherein said compound containing nitrogen is selected from the group consisting of pyridine, ethylenediamine, hexamethylenediamine, n-propylamine, monoethylamine, triethylamine, diethylamine, allylamine, acrylamide, aniline, acrylonitrile, 1,2,4-triazole, 5-amino-1H-tetrazole, propargylamine and acetonitrile.  
     
     
         21 . A measuring chip according to any one of  claims 1  to  15 , wherein a monomer material of said plasma polymerization layer is a compound containing sulfur.  
     
     
         22 . A measuring chip according to  claim 21 , wherein said compound containing sulfur is selected from the group consisting of dimethyl sulfide, methyl disulfide, ethanethiol, ethanedithiol, thiophen, mercaptoethanol and dithreitol.  
     
     
         23 . A measuring chip according to any one of  claims 1  to  15 , wherein said monomer material of a plasma polymerization layer is a compound containing a halogen.  
     
     
         24 . A measuring chip according to any one of  claims 1  to  15 , wherein said monomer material of a plasma polymerization layer comprises a compound having one or more groups selected from the group consisting of —COOH, —CHO, —SH, —NH 2 , —OH, ═NH, —CONH 2 , —NCO, —CH═CH 2 , ═C═O and  
       
         
           
           
               
               
           
         
       
     
     
         25 . A measuring chip according to any one of  claims 1  to  15 , wherein said monomer material of a plasma polymerization layer is a compound having —C≡CCH 2 OH.  
     
     
         26 . A measuring chip according to any one of  claims 1  to  15 , wherein said monomer material of a plasma polymerization layer is a carbohydrate compound comprising C and H.  
     
     
         27 . A measuring chip according to any one of  claims 1  to  15 , wherein said monomer material of a plasma polymerization layer is an organic metal compound.  
     
     
         28 . A measuring chip according to  claim 27 , wherein said organic metal compound is an organic silicon compound.  
     
     
         29 . A measuring chip according to  claim 28 , wherein said organic silicon compound is selected from the group consisting of tetramethylsilane, tetramethyldisiloxane, hexamethyldisiloxane, hexamethyldisilazane, hexamethylcyclotrisilazane, dimethylaminotrimethylsilane, trimethylvinylsilane, tetramethoxysilane, aminopropyltriethoxysilane, octadecyldiethoxymethylsilane, hexamethyldisilane, and divinyltetramethyldisiloxane.  
     
     
         30 . A measuring chip according to any one of  claims 1  to  15 , wherein plasma treatment is applied to said plasma polymerization layer with a polymeric or non-polymeric monomer.  
     
     
         31 . A measuring chip according to  claim 30  wherein said non-polymeric monomer material is selected from the group consisting of nitrogen, ammonium, hydrazine, hydrogensulfide, hydrogendisulfide, oxygen, hydrogen, water, halogen gas, and rare gas.  
     
     
         32 . A measuring chip according to any one of  claims 1  to  15 , wherein said monomer material of plasma polymerization layer is a mixture of two or more of the substances claimed in  claims 18  to  31 .  
     
     
         33 . A measuring chip for a surface plasmon resonance sensor comprising a metal layer, one or more plasma polymerization layers formed on said metal layer, and an immune protein or enzyme immobilized on the surface of said plasma polymerization layer, wherein said plasma polymerization layer comprises a monomer material selected from the group consisting of pyridine, triethylamine, diethylamine, allylamine, acrylamide, aniline, acrylonitrile, 1,2,4-triazole, 5-amino-1H-tetrazole, and acetonitrile.  
     
     
         34 . A measuring chip for a surface plasmon resonance sensor comprising a metal layer, one or more plasma polymerization layers formed on said metal layer, and an immune protein or enzyme immobilized on the surface of said plasma polymerization layer, wherein said plasma polymerization layer comprises a monomer material selected from the group consisting of the compounds claimed in any one of  claims 21  to  32 .  
     
     
         35 . A measuring chip according to  claim 33  or  34 , wherein said immune protein is an antibody.  
     
     
         36 . A measuring chip according to  claim 33  or  34 , wherein said immune protein is a Fab fragment of an antibody.  
     
     
         37 . A measuring chip according to  claim 33  or  34 , wherein said immune protein is a F(ab)2 fragment of an antibody.  
     
     
         38 . A measuring chip according to  claim 2  or  3 , wherein said nucleic acid is immobilized on said plasma polymerization layer through a cross-linking reagent or a condensation reagent.  
     
     
         39 . A measuring chip according to  claim 4  or  5 , wherein said non-immune protein is immobilized on said plasma polymerization layer through a cross-linking reagent or a condensation reagent.  
     
     
         40 . A measuring chip according to  claim 6  or  7 , wherein said immunoglobulin-binding protein is immobilized on said plasma polymerization layer through a cross-linking reagent or a condensation reagent.  
     
     
         41 . A measuring chip according to  claim 8  or  9 , wherein said sugar-binding protein is immobilized on said plasma polymerization layer through a cross-linking reagent or a condensation reagent.  
     
     
         42 . A measuring chip according to  claim 10 , wherein said sugar-recognizing sugar chain is immobilized on said plasma polymerization layer through a cross-linking reagent or a condensation reagent.  
     
     
         43 . A measuring chip according to  claim 11  or  12 , wherein said fatty acid or fatty acid ester is immobilized on said plasma polymerization layer through a cross-linking reagent or a condensation reagent.  
     
     
         44 . A measuring chip according to  claim 13 , wherein said polypeptide or oligopeptide is immobilized on said plasma polymerization layer through a cross-linking reagent or a condensation reagent.  
     
     
         45 . A measuring chip according to any one of  claims 38  to  44 , wherein said cross-linking reagent is one or more compounds selected from the group consisting of glutaraldehyde, periodic acid, N-succinimidyl-2-maleimidoacetic acid, N-succinimidyl-4-maleimidobutyric acid, N-succinimidyl-6-maleimidohexanic acid, N-succinimidyl-4-maleimidomethylcyclohexan-1-carboxylic acid, N-sulfosuccinimidyl-4-maleimidomethylcyclohexane-1-carboxylic acid, N-succinimidyl-4-maleimidomethylbanzoic acid, N-succinimidyl-3-maleimidobenzoic acid, N-sulfosuccinimidyl-3-maleimidobenzoic acid, N-succinimidyl-4-maleimidophenyl-4-butyric acid, N-sulfosuccinimidyl-4-maleimidophenyl-4-butyric acid, N,N′-oxydimethylene-dimaleimide, N,N′-o-phenylene-dimaleimide, N,N′-m-phenylene-dimaleimide, N,N′-p-phenylene-dimaleimide, N,N′-hexamethylene-dimaleimide, N-succinimidylmaleimidocarboxylic acid, N-succinimidyl-S-acetylmercaptoacetic acid, N-succinimidyl-3-(2-pyridyldithio)propionate, S-acetylmercaptosuccinic anhydride, methyl-3-(4′-dithiopyridyl)propionimidate, methyl-4-mercaptobutylimidate, methyl-3-mercaptopropionimidate, iminothiolene, o-carboxymethyl-hydroxylamine, azodiphenylpilmaleido, bis(sulfosuccinimidyl)sperate, 4,4′-diisothiocyano-2,2′-disulfonic acid stilbene, 4,4′-difluoro-3,3′-dinitrodiphenylsulfon, 1,5-difluoro-2,4-dinitrobenzene, p-phenylenediisothiocyanate, dimethyladipimidate, dimethylpimelimidate, dimethylsuberimidate, p-azidophenacylbromide, p-azidophenylglyoxal, N-hydroxysuccinimidyl-4-azidobenzoate, 4-fluoro-3-nitrophenylazide, methyl-4-azidobenzoimidate, N-5-azido-2-nitrobenzoyloxysuccinimide, N-succinimidyl-6-(4′-azido-2′-nitrophenylamino)hexanoate, 1,4-benzoquinone, N-succinimidyl-3-(2′-pyridyldithio)propionate, N-(4-maleimidobutyloxy)sulfosuccinimide sodium salt, N-(6-maleimidocaproyloxy)sulfosuccinimide sodium salt, N-(8-maleimidocaproyloxy)sulfosuccinimide sodium salt, N-(11-maleimidoundecanoyloxy)sulfosuccinimide sodium salt, N-[2-(1-piperazinyl)ethyl]maleimide bichloric acid, bisdiazobenzidine, hexamethylenediisocyanate, toluenediisocyanate, hexamethylenediisothiocyanate, N,N′-ethylenebismaleinimide, N,N′-polymethylenebisiodoacetamide, 2,4-dinitrobenzenesulfonate sodium salt, and diazo compounds; or said condensation reagent is one or more compounds selected from the group consisting of carbodiimide derivatives represented by RN═C═NR (or R′), N-hydroxysuccinimide, tri-n-butylamine, butyl chloroformate, and isobutyl isocyanide.  
     
     
         46 . A measuring chip according to any one of  claims 33  to  37 , wherein said immune protein or enzyme is immobilized on said plasma polymerization layer through a cross-linking reagent or a water-soluble condensation reagent.  
     
     
         47 . A measuring chip according to  claim 46 , wherein said cross-linking reagent is one or more compounds selected from the group consisting of glutaraldehyde, N-succinimidyl-4-maleimidomethylbanzoic acid, N-succinimidyl-3-maleimidobenzoic acid, N-succinimidyl-4-maleimidophenyl-4-butyric acid, N,N′-oxydimethylene-dimaleimide, N,N′-m-phenylene-dimaleimide, N,N′-p-phenylene-dimaleimide, N,N′-hexamethylene-dimaleimide, N-succinimidylmaleimidocarboxylic acid, N-succinimidyl-S-acetylmercaptoacetic acid, N-succinimidyl-3-(2-pyridyldithio)propionate, S-acetylmercaptosuccinic anhydride, methyl-3-(4′-dithiopyridyl)propionimidate, methyl-4-mercaptobutylimidate, methyl-3-mercaptopropionimidate, iminothiolene, o-carboxymethyl-hydroxylamine, azodiphenylpilmaleido, bis(sulfosuccinimidyl)sperate, 4,4′-diisothiocyano-2,2′-disulfonic acid stilbene, 4,4′-difluoro-3,3′-dinitrodiphenylsulfon, 1,5-difluoro-2,4-dinitrobenzene, p-phenylenediisothiocyanate, dimethyladipimidate, dimethylpimelimidate, dimethylsuberimidate, p-azidophenacylbromide, p-azidophenylglyoxal, N-hydroxysuccinimidyl-4-azidobenzoate, 4-fluoro-3-nitrophenylazide, methyl-4-azidobenzoimidate, N-5-azido-2-nitrobenzoyloxysuccinimide, N-succinimidyl-6-(4′-azido-2′-nitrophenylamino)hexanoate, 1,4-benzoquinone, N-succinimidyl-3-(2′-pyridyldithio)propionate, bisdiazobenzidine, hexamethylenediisocyanate, toluenediisocyanate, hexamethylenediisothiocyanate, N,N′-ethylenebismaleinimido, N,N′-polymethylenebisiodoacetoamide, and diazo compounds; or said condensation reagent is one or more compounds selected from the group consisting of carbodiimide derivatives represented by RN═C═NR (or R′), N-hydroxysuccinimide, tri-n-butylamine, butyl chloroformate, and isobutyl isocyanide.  
     
     
         48 . A measuring chip for a surface plasmon resonance sensor comprising a metal layer, a plasma polymerization layer formed on said metal layer, a substance immobilized on the surface of said plasma polymerization layer, and an additional plasma polymerization layer or plasma-treated layer formed on said plasma polymerization layer.  
     
     
         49 . A measuring chip according to  claim 48 , wherein said substance to be immobilized is selected from the group consisting of a nucleic acid, a non-immune protein, an immunoglobulin-binding protein, a sugar-binding protein, a sugar-recognizing sugar chain, a fatty acid or a fatty acid ester, a polypeptide or oligopeptide having ligand-binding activity, an immune protein, and an enzyme.  
     
     
         50 . A measuring chip for a surface plasmon resonance sensor, comprising a metal layer, one or more plasma polymerization layers formed on said metal layer, and a substance immobilized on said plasma polymerization layer through a hydrophobic bond.  
     
     
         51 . A measuring chip according to  claim 50 , wherein said substance to be immobilized is selected from the group consisting of a nucleic acid, a non-immune protein, an immunoglobulin-binding protein, a sugar-binding protein, a sugar-recognizing sugar chain, a fatty acid or a fatty acid ester, a polypeptide or oligopeptide having ligand-binding activity, an immune protein, and an enzyme.  
     
     
         52 . A method for producing a measuring chip for a surface plasmon resonance sensor comprising the steps of forming a metal layer on an optically transparent substrate, forming one or more plasma polymerization layers on said metal layer, and then immobilizing a nucleic acid on the surface of said plasma polymerization layer.  
     
     
         53 . A method for producing a measuring chip for a surface plasmon resonance sensor comprising the steps of forming a metal layer on an optically transparent substrate, forming one or more plasma polymerization layers on said metal layer, and then immobilizing a non-immune protein on the surface of said plasma polymerization layer.  
     
     
         54 . A method for producing a measuring chip for a surface plasmon resonance sensor comprising the steps of forming a metal layer on an optically transparent substrate, forming one or more plasma polymerization layers on said metal layer, and then immobilizing an immunoglobulin-binding protein on the surface of said plasma polymerization layer.  
     
     
         55 . A method for producing a measuring chip for a surface plasmon resonance sensor comprising the steps of forming a metal layer on an optically transparent substrate, forming one or more plasma polymerization layers on said metal layer, and then immobilizing a sugar-binding protein on the surface of said plasma polymerization layer.  
     
     
         56 . A method for producing a measuring chip for a surface plasmon resonance sensor comprising the steps of forming a metal layer on an optically transparent substrate, forming one or more plasma polymerization layers on said metal layer, and then immobilizing a sugar-recognizing sugar chain on the surface of said plasma polymerization layer.  
     
     
         57 . A method for producing a measuring chip for a surface plasmon resonance sensor comprising the steps of forming a metal layer on an optically transparent substrate, forming one or more plasma polymerization layers on said metal layer, and then immobilizing a fatty acid or fatty acid ester on the surface of said plasma polymerization layer.  
     
     
         58 . A method for producing a measuring chip for a surface plasmon resonance sensor comprising the steps of forming a metal layer on an optically transparent substrate, forming one or more plasma polymerization layers on said metal layer, and then immobilizing a polypeptide or oligopeptide having a ligand binding capability on the surface of said plasma polymerization layer.  
     
     
         59 . A method according to any one of  claims 52  to  58 , wherein the plasma polymerization layer is formed by a plasma-treatment using a monomer material claimed in any one of  claims 18  to  32 .  
     
     
         60 . A method for producing a measuring chip for a surface plasmon resonance sensor comprising the steps of forming a metal layer on an optically transparent substrate, forming one or more plasma polymerization layers on said metal layer by a plasma-treatment using a monomer material claimed in any one of  claims 18  to  32 , and then immobilizing an immune protein or enzyme on the surface of said plasma polymerization layer.  
     
     
         61 . A measuring cell for a surface plasmon resonance sensor comprising a measuring chip according to any one of  claims 1  to  51 .  
     
     
         62 . A measuring cell according to  claim 61 , wherein said chip is optically analyzed.  
     
     
         63 . A surface plasmon resonance biosensor comprising a measuring chip according to any one of  claims 1  to  51  or a measuring cell according to  claim 61  or  62 .