Biosensor

Abstract

It is an object of the present invention to provide a sensor, which is capable of measuring, quickly and in high accuracy, concentration of neutral fat from a sample such as a biological sample or the like, without executing pretreatment of the sample. This object is attained by a biosensor for measuring concentration of neutral fat, based on value of current flowing in the electrode system, having: an insulating substrate; an electrode system having a working electrode and a counter electrode, formed onto the insulating substrate; and a reaction layer having a lipoprotein lipase, a glycerol dehydrogenase and an electron mediator, formed at the upper part or the vicinity of the electrode system.

1 . A biosensor for measuring concentration of neutral fat, based on value of current flowing in said electrode system, comprising:
 an insulating substrate;   an electrode system having a working electrode and a counter electrode, formed onto said insulating substrate; and   a reaction layer having a lipoprotein lipase, a glycerol dehydrogenase and an electron mediator, formed at the upper part or the vicinity of said electrode system.   
   
   
       2 . The biosensor according to  claim 1 , wherein said electrode system further comprises a reference electrode. 
   
   
       3 . The biosensor according to  claim 1 , wherein said glycerol dehydrogenase is a coenzyme dependent type. 
   
   
       4 . The biosensor according to  claim 3 , wherein said coenzyme is pyrrolo-quinoline quinone or flavin adenine dinucleotide. 
   
   
       5 . The biosensor according to  claim 1 , wherein said electron mediator is one kind or two or more kinds selected from the group consisting of a ferricyanide ion, p-benzoquinone and a p-benzoquinone derivative, phenazine methosulfate and a phenazine methosulfate derivative, methylene blue, thionin, indigocarmine, gallocyanin, safranin, ferrocene and a derivative thereof, α-naphthoquinone and an α-naphthoquinone derivative. 
   
   
       6 . The biosensor according to  claim 1 , wherein said reaction layer further comprises a surfactant and a pH buffering agent. 
   
   
       7 . The biosensor according to  claim 6 , wherein said surfactant is a nonionic surfactant. 
   
   
       8 . The biosensor according to  claim 6 , wherein pH of said pH buffering agent is 6 to 9. 
   
   
       9 . The biosensor according to  claim 1 , wherein said reaction layer further comprises a hydrophilic polymer. 
   
   
       10 . The biosensor according to  claim 9 , wherein said hydrophilic polymer is one kind or two or more kinds selected from the group consisting of tannic acid, pectin, casein, carrageenan, furcellaran, pullulan, collagen, chitin, chitosan, sodium chondroitin sulfate, lignin sulfonic acid, and derivatives thereof. 
   
   
       11 . The biosensor according to  claim 9 , wherein said reaction layer comprises:
 a first reaction layer having said electron mediator and said hydrophilic polymer, but substantially not having said glycerol dehydrogenase; and   a second reaction layer formed onto the upper layer of said first reaction layer, and having said glycerol dehydrogenase and said hydrophilic polymer, but substantially not having said electron mediator.   
   
   
       12 . The biosensor according to  claim 11 , wherein a third reaction layer consisting essentially of said hydrophilic polymer is formed between said first reaction layer and said second reaction layer. 
   
   
       13 . The biosensor according to  claim 1 , wherein an insulating layer having a nearly rectangular-shape opening part is formed at the upper layer of said electrode system, characterized in that exposure area from said opening part of said counter electrode is 1.2 to 3.0 times exposure area from said opening part of said working electrode. 
   
   
       14 . The biosensor according to  claim 1 , further comprising a lead part and a connector part for making said electrode system electrically conducted to outside of the biosensor, wherein a material configuring said electrode system is a material of carbon, as a main component, with a surface resistance resistivity value of equal to or lower than 100Ω/□, in a thickness of 10 μm, and a material configuring said lead part and said connector part is a material of metal, as a main component, with a surface resistance resistivity value of equal to or lower than 50Ω/□, in a thickness of 10 μm. 
   
   
       15 . The biosensor according to  claim 14 , wherein said metal is one kind or two or more kinds of metals selected from the group consisting of silver, gold, platinum and palladium. 
   
   
       16 . The biosensor according to  claim 3 , wherein said glycerol dehydrogenase is a modified glycerol dehydrogenase chemically modified with a bivalent cross-linking reagent, in the presence of a surfactant. 
   
   
       17 . The biosensor according to  claim 16 , wherein said glycerol dehydrogenase is derived from bacteria belonging to Gluconobacter. 
   
   
       18 . The biosensor according to  claim 2 , wherein said glycerol dehydrogenase is a coenzyme dependent type. 
   
   
       19 . The biosensor according to  claim 2 , wherein said electron mediator is one kind or two or more kinds selected from the group consisting of a ferricyanide ion, p-benzoquinone and a p-benzoquinone derivative, phenazine methosulfate and a phenazine methosulfate derivative, methylene blue, thionin, indigocarmine, gallocyanin, safranin, ferrocene and a derivative thereof, α-naphthoquinone and an α-naphthoquinone derivative. 
   
   
       20 . The biosensor according to  claim 3 , wherein said electron mediator is one kind or two or more kinds selected from the group consisting of a ferricyanide ion, p-benzoquinone and a p-benzoquinone derivative, phenazine methosulfate and a phenazine methosulfate derivative, methylene blue, thionin, indigocarmine, gallocyanin, safranin, ferrocene and a derivative thereof, α-naphthoquinone and an α-naphthoquinone derivative. 
   
   
       21 . The biosensor according to  claim 4 , wherein said electron mediator is one kind or two or more kinds selected from the group consisting of a ferricyanide ion, p-benzoquinone and a p-benzoquinone derivative, phenazine methosulfate and a phenazine methosulfate derivative, methylene blue, thionin, indigocarmine, gallocyanin, safranin, ferrocene and a derivative thereof, α-naphthoquinone and an α-naphthoquinone derivative. 
   
   
       22 . The biosensor according to  claim 18 , wherein said electron mediator is one kind or two or more kinds selected from the group consisting of a ferricyanide ion, p-benzoquinone and a p-benzoquinone derivative, phenazine methosulfate and a phenazine methosulfate derivative, methylene blue, thionin, indigocarmine, gallocyanin, safranin, ferrocene and a derivative thereof, α-naphthoquinone and an α-naphthoquinone derivative.