KR101738207B1 - Cross-linkable nanostructured organometallic polymers for enzymatic biofuel cell and biosensor applications - Google Patents

Abstract

The present invention relates to a bioelectrode containing a crosslinkable organometallic polymer and a method of manufacturing the same, and more particularly, to an electrode having nanostructure of an organometallic polymer controlled for use in a biofuel cell, a biosensor, .
The electrode according to the present invention includes an organic metal and includes a self-assembling block copolymer and an enzyme. The double continuous structure shows the maximum current value at the current density of the electrode manufactured at the same glucose concentration , Exhibits good sensitivity even at low glucose concentrations and provides applications for biofuel cells and biosensors.

Description

Technical Field [0001] The present invention relates to an electrode including a self-assembling polymer containing an organic metal,

The present invention relates to a bioelectrode containing a crosslinkable organometallic polymer and a method of manufacturing the same, and more particularly, to an electrode having nanostructure of an organometallic polymer controlled for use in a biofuel cell, a biosensor, .

Effective Enzymes The development of biofuel cells has been an important research topic over the past several decades as an energy source for biomedical devices, microchip systems, and portable electronic devices.

BACKGROUND ART Biofuel cells that produce electricity using biomass such as alcohols and glucose are attracting much attention due to their environmental friendliness and permanent sustainability. However, such a biofuel cell has a disadvantage in that it has a smaller power density than other energy sources.

The main reason for this is that the redox active site in the structure of the enzyme is hidden inside the structure due to the stability of the enzyme, which causes the oxidation-reduction reaction and the electron transfer rate to decrease do. Low electron transfer and slow oxidation-reduction reactions are impeding the building of energy systems for the miniaturization of biomedical devices.

Therefore, various studies have been made to increase the power density of the biofuel cell. A method called direct electron transfer is one of the most widely studied methods to increase the power density, and is to control the catalytic active site of the enzyme within the electron tunneling distance of the electrode surface. For this reason, electronic intermediates are attracting much attention as a role in assisting electron transfer in the transport of the enzyme from the catalytically active site to the current collector. Therefore, studies on various materials with the capability of electronic intermediation have been actively conducted in recent years and many reports have been made. These materials include nano-carbon based materials, oxidation-reduction active polymers, cofactor relays, and metal nanoparticles.

In order to improve the stability of the electrode and the current density in the presence of the electron mediator, enzyme fixation at the electrode is essential. For this reason, a considerable number of studies on electronic mediators have been conducted in relation to the electrical coupling of enzymes on the electrode surfaces to which electronic mediators are bound.

From this 'production' perspective, carbon nanotubes have received much attention due to their wide specific surface area and good electrochemical properties. However, such a carbon nanotube has a disadvantage in that it is difficult to prevent the aggregation of carbon nanotubes when a chemical modification process for arranging each carbon nanotube is not performed in manufacturing a large-area electrode.

Studies on oxidation-reduction active polymers have also been carried out. Many types of organometallic polymers have been known to be effective for transferring electrons through oxidation-reduction reactions. In this case, it is known that the concentration of the oxidation-reduction moiety, the oxidation-reduction potential, and the control of many chemical linkers and functionalization of the oxidation-reduction polymer play an important role in controlling electron transfer rate.

On the other hand, PCT / JP2003 / 009480 filed by Sony discloses methods for producing an optimal fuel cell using various enzymes or complex enzymes.

However, there is a lack of research to improve the performance of electron transfer by controlling the shape of the oxidation-reduction polymer. Therefore, there is a demand for a more efficient and stable new structure of the oxidation-reduction polymer.

A problem to be solved by the present invention is to provide a redox polymer of a novel structure having high current density and stability.

Another object of the present invention is to provide a novel electrode having a high current density and stability, which is composed of an enzyme and an oxidation-reduction polymer.

Another object of the present invention is to provide a method for controlling the shape of an oxidation-reduction polymer.

Another object of the present invention is to provide a new type of biofuel cell.

In order to solve the above problems, the present invention comprises an electrode comprising a self-assembling block copolymer containing an organic metal and an enzyme.

The present invention also comprises a fuel cell comprising a self-assembling block copolymer comprising an organometallic and an electrode comprising an enzyme.

In the present invention, the enzyme produces electrons using biomass, and the produced electrons can produce electrons transferred through the organic metal. The enzyme can be produced by a variety of known enzymes according to the mechanism of producing electrons Can be used. For example, it is preferable that the enzyme is selected by using the optimum enzyme according to the fuel. For example, glucose dehydrogenase, a series of enzymes of the electron transport system, ATP synthase, enzymes involved in glucose metabolism (e.g., hexokinase, glucose phosphate isomerase, phosphooroktiana, orthoose 2 Phosphoric acid aldorase,

L-lactic acid dihydrogenase, D-lactic acid dihydrogenase, glyceraldehyde dehydrogenase, glyceraldehyde dehydrogenase, phosphoglyceromutase, phosphopyrimidine dehydratase, pyruvate kinase, Acyltransferase, azine, pyruvic acid dihydrogenase, citric acid synthase, aconitase, isocitrate dihydrogenase, 2-oxoglutarate dihydrogenase, succinyl-CoA synthetase, zucaric acid dihydrogenase, Azelaic acid, malonic acid dihydrogenase, etc.), and glucose can be combined with glucose oxidase.

Examples of the fuel cell of the present invention include, in addition to saccharides such as glucose, organic acids such as ethanol and the like, alcohols such as alcohols, fatty acids, proteins, and intermediate products of glucose metabolism (glucose-6-phosphate, 3-phosphoglycerinic acid, 2-phosphoglycerinic acid, phosphoenolpyruvic acid, pyruvic acid, acetyl-CoA, pyruvic acid, isoamylase, Citric acid, cis-aconitic acid, iso-citric acid, oxalo succinic acid, 2-oxoglutaric acid, succinyl-CoA, succinic acid, malic acid, L-malic acid and oxaloacetic acid) . Among them, glucose, ethanol, an intermediate product of glucose metabolism and the like can be used. In particular, glucose is a preferable fuel because it is extremely easy to handle.

In the present invention, the organic metal is not particularly limited when it is a conductive metal capable of transferring electrons generated by the enzyme.

In the present invention, the block copolymer is a block copolymer at least partially crosslinked, and preferably includes a crosslinkable block. The crosslinkable block may be a block containing at least one unsaturated group such as a double bond, and may be cross-linked by a metal such as Os (Os).

In the present invention, the block copolymer may have an amorphous double continuous structure, a crystalline form of nanowire, or a nanoparticle form through metal self-assembly process, preferably an enzyme mixed with a crosslinked block copolymer It is preferable to have an amorphous double continuous structure so as to increase the contact of the semiconductor wafer. The amorphous double continuous structure means a shape as shown in the drawing, and several metal domains can be expressed in the form of interconnected by a band.

In the present invention, the form of the block copolymer may vary depending on the molecular weight, the kind of the solvent, and the affinity with the solvent.

In a preferred embodiment of the present invention, the block copolymer may be represented by the following general formula, preferably poly (ferrocenyldimethylsilane-b-isoprene).

Wherein x and y are degrees of polymerization and R1 and R2 are independently hydrogen or alkyl, acyl, or alkoxy having 1 to 30 carbon atoms and the molecular weight of each block is 0.1-500 kg / mol, preferably 1-100 kg / mol.

In the present invention, the enzyme may be prepared in the form of a solution together with the self-assembling block copolymer, and then coated to form a coating film. The coating layer preferably has a thickness of 1-50 micrometers, and the enzyme is preferably 1-50 wt%, preferably about 5-45 wt%, and most preferably 30 wt% of the entire coating film .

In one aspect, the present invention provides block copolymers that are self assembled in the form of an amorphous binary continuous structure or nanoparticles comprising a conductive organometallic block.

According to another aspect of the present invention, there is provided a biofuel cell characterized in that an electric current is produced using an electrode comprising a self-assembling block copolymer containing an organic metal and an enzyme. The biofuel cell is a fuel cell that reacts glucose or an alcohol with an enzyme to obtain an electric current.

According to another aspect of the present invention, there is provided a method for producing an electric current using a self-assembling block copolymer comprising an organic metal and an electrode comprising an enzyme, Lt; / RTI >

The present invention relates to the electrochemical characterization of enzymes integrated with nanometer-sized self-assembled oxidation-reduction polymers.

In the present invention, an enzyme used as a preferred embodiment is glucose oxidase (GOx) and an organic metal block copolymer including poly (ferrocenyldimethylsilane) is used as an electron mediator. In the present invention, the synthesis of a block copolymer containing a ferrocene (Fc) and the natural self-assembling portion of the polymer were used to increase the electron transfer rate between the flavin adenine dinucleotide (FAD) it is possible to control the molecular weight and the solvent fraction of the ferrocenyldimethylsilane-b-isoprene (PFDMS-b-PI) block copolymer and the PFDMS-b-PI block copolymer to form bicontinuous structures, nanowires, And the large catalytic current characteristics are changed by the change of the PFDMS-b-PI nanostructure. When PFDMS-b-PI of double continuous structure is used as an electron mediator, current density is improved by 2 ~ 50 times than that of other forms. In addition, when the PI chain using osmium (Os) bond is cross-linked, the stability of the electrode manufactured under physiological conditions is increased.

In one embodiment, the present invention can be used as a glucose biosensor using an electrode, and can be applied to bio-micro devices inserted into the body.

In the present invention, a functional electrode composed of a nanostructured organometallic block copolymer capable of cross-linking with an enzyme was prepared, and the electron transfer rate of the enzyme was improved, and the stability of the electrode was improved through cross-linking. In addition, a method of determining the electrochemical properties of the enzyme biofuel cell by controlling the shape of the oxidation-reduction polymer-based electronic mediator through the present invention has been proposed. The double continuous structure shows the maximum current value at the current density of the electrode made at the same glucose concentration and shows good sensitivity even at low glucose concentration and provides the application to the biofuel cell and the biosensor.

1 is a TEM image showing the structure of a PFDMS-b-PI block copolymer in different mixed solvents.
FIG. 2 is a graph showing an electrode made of PFDMS-b-PI _OS and GOx having a nanowire structure using FIB-TEM.
FIG. 3 shows the structure of the electrode including the PFDMS-b-PI _OS having a double continuous structure, which was observed using a scanning electron microscope.
FIG. 4 is a graph showing a measurement of cyclic voltammetry using the glucose concentration in order to confirm how the effect of the electron mediator varies depending on the polymer structure in the electrode.
5 is a graph showing the current measured while adding glucose in a physiological environment.
Figure 6 shows the process of making a functional electrode composed of GOx and PFDMS-b-PI intermediates based on the nanowire structure of the polymer.

Example

Synthesis of 1,1'-dimethylsilylferrocenophane monomer

The 1,1'-dimethylsilylferrocenophane monomer containing ferrocene was synthesized by coupling silane with lithiated ferrocene. This method was introduced by the Manner team, which was introduced as a reference in the present invention. The synthesized monomers were purified by sublimation and recrystallization under vacuum.

Synthesis of PFDMS-b-PI

PFDMS-b-PI block copolymers with different molecular weights were synthesized through sequential anionic polymerization. PI precursor synthesis was carried out using purified benzene as solvent and s-butyllithium as initiator. After the PI chain of the desired molecular weight is synthesized, the previously measured mass of 1,1'-dimethylsilylferrocenophane monomer is added to the reactor in the glove box. The reactor is then turned back into a vacuum line and the gas is completely removed. A small amount of purified THF solvent is distilled in the reactor to increase the polymerization rate of the 1,1'-dimethylsilylferrocenophane monomer. After the polymerization is completed, the color of the reaction solution changes from red to brown. Polymerization proceeds for a minimum of 6 hours and is then terminated by isopropanol. In the final reaction, the color of the solution changes from brown to red. The PFDMS-b-PI block copolymer was precipitated with hexane. The polydispersity indices of the synthesized block copolymers were measured to be 1.08.

Electrode production

Glucose oxidase enzyme (~ 200 U / mg) obtained from Aspergillus niger was purchased from Sigma-Aldrich. To prepare a functional electrode composed of GOx and a block copolymer, a 1 wt% PFDMS-b-PI solution containing 60 μM of GOx aqueous solution and various solvents was prepared. Then, each solvent is uniformly mixed and directly sprayed onto the porous carbon electrode through a drop coating method. After removing all solvents with a vacuum oven, the electrode is exposed to OsO4 vapor for 3 hours. The electrode is then washed with distilled water (Milli-Q Water) and stored in PBS buffer before use.

Electrochemical experiment

Cyclic voltammetry (CV) experiments were recorded on an EG & G PAR 273 instrument and measured with a three electrode system using platinum gauze as a counter electrode and a silver / silver chloride electrode as a reference electrode. All potentials given here were obtained on the basis of silver / chloride reference electrodes. The working electrode is a porous carbon electrode prepared according to the present invention. All experiments were performed in PBS buffer (pH 7.4) at room temperature, atmospheric pressure, and air. The control experiment with only the PBS buffer solution yielded a characteristic flat CV result. All electrochemical experiments were repeated with more than 20 electrodes and results were shown for the most typical scan rate of 20 mV / s. For stability of the results, the results measured from the first cycle were deleted and recorded from the results of the next cycle.

Morphological feature analysis

The surface structure of the fabricated electrode was observed using an atomic force microscope (AFM) in a tapping mode. All measurements were performed within a phase difference of 10 degrees. Transmission Electron Microscopy (TEM) Samples were prepared by drop coating method using a PFDMS-b-PI block copolymer having a concentration of 1 wt% in a mixed solvent. Before the measurement, the block copolymer was stained with OsO4 vapor to increase the electron contrast of the Fc portion and the PI chain portion. The microscope used was a Zeiss LIBRA 200 FE microscope and measured with an Omega energy filter at an energy of 200 kV at -160 ° C. The iron element mapping was measured using the three-window method using the iron energy level of 709 eV. A TEM sample etched with a focused ion beam was fabricated using a cross-sectional FIB-TEM FEI Strata 235 Dual-beam FIB and 30 kV energy was used and exposed to RuO4 vapor for 10 hours for surface protection. Imaging of the etched samples was measured with an energy of 200 kV using a JEOL 3010 microscope.

Conductivity measurement

The conductivity of the PFDMS-b-PI block copolymer was measured by AC impedance spectroscopy in a glove box. A through-plane conductivity of 1.15 cm x 2 cm was used as a working electrode and a counter electrode. The resulting collection was collected over a frequency range of 10-100,000 Hz using a 1260 Solatron impedance analyzer.

Characterization of PFDMS-b-PI block copolymer

The block copolymers PFDMS-b-PI containing diene chains and ferrocene (Fc), which are reactive with osmium, were synthesized through sequential anionic polymerization. Two different PFDMS-b-PIs were obtained and their molecular weights were 10.4-8.0 kg / mol and 69.0-92.0 kg / mol. The chemical structure of the PFDMS-b-PI block copolymer in Formula (1) is shown, wherein x and y each represent the degree of polymerization of each chain. The PFDMS chain containing Fc in the block copolymer has the capability of oxidation-reduction reaction. The electrochemical properties of Fc in the polymer are similar to those of small Fc molecules.

        1B-e shows TEM images of the structure of PFDMS-b-PI block copolymer in different mixed solvents. It can be confirmed that the PFDMS-b-PI block copolymer has different nanostructures depending on the molecular weight and the fraction of the mixed solvent. For example, in the case of a small molecular weight PFDMS-b-PI (10.4-8.0 kg / mol), it has a double continuous structure with a size of 200 nm in a mixed solution of toluene and hexane (20/80 vol% In FIG. 1C, it can be seen that the portion darkened in FIG. 1B is rich in iron using iron element mapping. On the other hand, in the mixed solution of THF and hexane (20/80 vol.%), Another structure of several micron-length nanowires was observed, and the diameter of the nanowires was observed to be 20 nm. Though not to be bound by theory, this can be explained by the solubility parameter, in which the dissolution parameter of PFDMS (18.6 MPa < 1/2 >) has a value similar to the solubility parameter values of toluene (18.3 MPa & MPa < 1/2 >) has a value close to that of hexane (14.9 MPa < 1/2 >). In addition, it can be confirmed that the crystalline nature of (toluene and THF) PFDMS behaves differently in selecting the solvent.

On the other hand, when a solution was prepared by using a large molecular weight PFDMS-b-PI (69.0-92.0 kg / mol) in a mixed solution of THF and hexane (20/80 vol%), uniform nanoparticles of 60 nm in size were obtained. We also confirmed the structure of nano-wires with a short diameter of 40 nm when the ratio of THF to hexane in the solution (25/75 vol%) was not uniform. This means that the nanoparticle structure is stable only in a very narrow THF / hexane fraction.

This nanostructured PFDMS-b-PI organometallic block copolymer is used to transfer electrons from the GOx / glucose oxidation-reduction reaction to the electrode. Figure 6 shows a functional electrode composed of GOx and PFDMS-b-PI intermediates. . For ease of illustration, FIG. 6 is based on the nanowire structure of the polymer. A homogeneous GOx solution and a PFDMS-b-PI block copolymer solution were prepared and dropped on a porous carbon electrode. The thickness of the coated film varied from 1 to 50 micrometers and we measured the maximum current density to find the optimal thickness. The maximum catalytic current was measured at a GOx concentration of 30 wt%, and precipitation occurred when the enzyme concentration exceeded 50 wt%.

In the present invention, the PI chain used as the auxiliary polymer provides a cross-linking position of the diene group. When the OsO4 vapor is exposed to the OsO4 vapor for more than 3 hours for chemical cross-linking, Os of OsO4 forms a covalent bond with the hydrocarbon, . Hereinafter, the prepared intermediate material polymer will be referred to as PFDMS-b-PI _OS . Os After staining, wash with distilled water to remove unfixed GOx. Electrodes not coated with Os are very unstable under physiological conditions. In the final step, an oxidation potential was applied to activate the Fc portion. After activation, the Fc moiety becomes more strongly bound to the negative charge of GOx by positively charging.

Structural Analysis of Fabricated Electrode

The structures of the fabricated electrodes were observed using FIB, TEM, and SEM. FIG. 2 is a graph showing an electrode made of PFDMS-b-PI _OS and GOx having a nanowire structure using FIB-TEM. The FIB-TEM experiment was performed after ruthenium (Ru) treatment on the electrode surface to minimize damage caused by electron beam during the experiment. As shown in FIG. 2, it can be seen that the PFDMS-b-PI _OS mediator and the GOx enzyme on the electrode have an intermixing structure and an interpenetration structure. The thickness of this layer is 100 to 200 nanometers, which corresponds to the lower portion of the substrate. As shown in the enlarged drawing of the inside figure, it can be confirmed that the nanowire structure having a diameter of 20 nm is seen, and it is confirmed that the GOx enzyme which has a size of 40 to 80 nanometers is observed in the internal mixing structure. This indicates that the experimental conditions are very unstable with respect to the electron beam despite the cryogenic temperature (-160 ^° C) and the low dose rate (9.6 e- / Å ² ). A representative example is shown in a white inner illustration, represented by another context.

The structures of the electrodes including the PFDMS-b-PI _OS with the double continuous structure were observed using a scanning electron microscope. As shown in FIG. 3A, the upper part of the manufactured electrode shows a state of the double continuous structure shown in FIG. 1B. The transverse cross-section of the electrode shows that it is porous enough to allow glucose and enzyme to meet even though there is a protective film that fixes GOx. In addition, the contact surface between the current collector and GOx / PFDMS-b-PI _{OS was} cut in liquid nitrogen to determine the shape of the bottom of the electrode. As can be seen from FIG. 3B, it can be seen that the double continuous structure is continuously observed, indicating that the PFDMS-b-PI _OS polymer is completely present on the electrode surface and the electrode. However, since the size of a polymer having a double continuous structure is larger than that of a general electrode material (80-120 nm), it has been difficult to confirm the structure clearly in the FIB-TEM at a cross section (200 nm).

Glucose oxidation to a functional electrode composed of GOx / PFDMS-b-PI _OS mediator

PFDMS-b-PI In order to investigate the efficiency of the GOx fixed and arranged by the network of the _OS , the characteristics of the electrode were measured by the cyclic voltammetry (CV). The concentration of Fc in the electrode was fixed at 2mM and measured at a low scan rate of 20mV / s. As shown in FIG. 4a, when GOx was not present, the electrochemical potentials of Ag / AgCl electrode based Fc were 420mV and 550mV And it was confirmed that it shows a clear current change. This change is valid considering that the well known standard reduction potential of Fc is E ^o = 400 mV. However, the electrode mixed with GOx was able to detect other oxidation-reduction changes. It can be clearly seen that the reduction peak of 420 mV, which was previously located at 390, -80 mV, disappears at the positions of the oxidation and reduction peaks. The change in the oxidation peak from 550 to 390 mV is also thought to be due to the change in solvation of Fc due to the polar environment provided by GOx. The disappearance of the 420mV reduction peak is believed to be due to the transfer of electrons to the GO (FAD) material in the Fc + state and the Fc is reoxidized and converted to the Fc + state.

GO (FADH ₂ ) + 2Fc ⁺ -> GO (FAD) + 2Fc ⁰ + 2H ⁺

The standard reduction potential of FAD is well known to be -180 mV, and the electrochemical change of these electrodes is believed that GOx successfully binds to the PFDMS-b-PI _OS polymer network and effectively interacts with the Fc region to obtain enzymatic activity. Also, we have not seen any reaction to the oxidation-reduction of osmium here (Os (III) / Os (II)), suggesting that the Os moiety is sensitive to local concentration.

In order to investigate the effect of electron mediation on the polymer structure, the cyclic voltammetry was measured using glucose concentration. The PFDMS-b-PI _OS polymer having a nanowire structure is used as an electron mediator, as shown in FIG. 4B, and glucose was added to the PBS buffer solution. As can be seen, the addition of only 1.3mM glucose increased the peak of the oxidation-reduction current by 40%. As the concentration of glucose increased, the peak of oxidation and reduction current also steadily increased and the glucose concentration continued to be 60 mM. The internal figure is a graph showing the reduction current value of -80 mV according to the glucose concentration, and the AFM image shows the surface structure of the manufactured electrode.

PFDMS-b-PI _OS Oxidation-reduction process of electrode shows big change depending on structure of intermediates. When the structure of the PFDMS-b-PI _OS polymer is changed into a double continuous structure as shown in FIG. 4c, a redox graph similar to that of a nanowire can be obtained. However, a peak current of about twice the magnitude of the peak current flows and is expressed as 550 μA / cm ² at a position of -150 mV at 60 mM glucose concentration. This phenomenon can be explained by the fact that PFDMS with double continuous structure is in contact with GOx trapped with large surface area to increase the electron transfer rate. Also, it can be seen that the peak current is in a very stable state due to the reduction of 2% in the peak structure regardless of the structure of the polymer even under the oxidation-reduction cycle of 75 times or more.

FIG. 4d shows the current density of the electrode mixed with GOx and PFDMS-b-PI _OS polymer according to the shape of the polymer. Specific CV results with 8mM glucose concentration are shown, which is similar to glucose concentration in our blood. Comparing the nanowires, nanoparticles, and the double continuous structure, PFDMS, which has a double continuous structure, shows the best oxidation-reduction reaction with GOx, which shows that the reduction peak of double- Providing a sufficient basis for moving to the left of the peak. Interestingly, in the case of a polymer having a nanoparticle structure as shown in FIG. 3D, the oxidation-reduction reaction is negligibly small because the PFDMS polymer is in an isolated circular form and is difficult to contact with GOx or glucose Can be explained. Therefore, it was confirmed that the ability of the functional electrode to determine the electron density of the system is greatly determined by the structure of the redox block copolymer through these experiments.

The electrode current was measured by cyclic voltammetry using an electrode fabricated using only a PFDMS-b-PI _OS block copolymer having a double continuous structure. As a result of the measurement, when compared with the PFDMS-b-PI _OS block copolymer having the nanowire structure made of the same thickness by using the same molecular weight, the double continuous structure was able to obtain a current twice as large. (Fig. 4S) This shows that the PFDMS-b-PI _OS block copolymer having a double continuous structure is oxidized twice more effectively than the PFDMS-b-PI _OS block copolymer having a nanowire structure due to its wide contact area. According to the efficiency of the oxidation of iron, a completely different conductivity value can be obtained. In order to clarify the effect of this structure more clearly, we measured the conductivity of the horizontal plane at room temperature and confirmed how the electron transfer occurred in GOx and substrate. Interestingly, the conductivity of the PFDMS-b-PI _OS block copolymer with double continuous structure was 1.34 x 10 ^-5 S / cm, and the conductivity value of PFDMS-b-PI _OS block copolymer was 4.03 x 10 Conductivity values of ^-6 S / cm were obtained.

The biosensing effect of the functional electrode according to the present invention was examined. In order to investigate the biocatalytic activity of the electrodes, experiments were carried out to inject various concentrations of glucose into the three electrode system. Since the concentration of physiological glucose in blood is between 5 and 10 mM, it is very important to show a large electrical signal even at low glucose concentration around 10 mM. Electrodes fabricated using a double continuous PFDMS-b-PI _OS block copolymer were sensitive to very low amounts of GOx (10 μl). The measurement of the current was carried out by measuring a reduction current peak value of -0.2 V based on the silver / silver chloride reference electrode. As shown in FIG. 5, by adding glucose in a physiological environment, the graph of the stepwise change shows that the Fc portion of the enzyme can be stably proportional to glucose concentration, I could confirm. Especially, at 1.3mM, which is very low concentration, it shows fast and accurate current change from 7 μA to 12 μA. Also, it was confirmed that the functional electrode of the present invention based on PFDMS-b-PI _OS block copolymer was in a very stable state while continuously measuring the glucose concentration for about 35 minutes while regularly changing the glucose concentration.

Claims (20)

A self-assembling block copolymer comprising an organic metal and an enzyme,
Wherein the self-assembling block copolymer including the organic metal is self-assembled into an amorphous bicontinuous structure, and the cross-linkable block including the double bond is crosslinked to fix the enzyme. delete delete delete The electrode according to claim 1, wherein the block copolymer is poly (ferrocenyldimethylsilane-b-isoprene). The electrode of claim 1, wherein the electrode comprises a self-assembling block copolymer comprising an organometallic and the enzyme forms a coating layer of 1-50 micrometers. 7. The electrode of claim 6, wherein the coating further comprises a porous carbon layer at the bottom. 8. The electrode of claim 7, wherein the enzyme in the coating layer is 1-50 wt%. The electrode according to claim 8, wherein the coating layer has an enzyme content of 30 wt%. delete delete The electrode according to claim 1, wherein the block copolymer is represented by the following formula (1).

Wherein x and y are degrees of polymerization and R1 and R2 are independently hydrogen or alkyl, acyl, alkoxy of 1-30 carbon atoms and the molecular weight of each block is 0.1-500 kg / mol. delete The biofuel cell according to claim 1, wherein the electrode is used to produce a current. A biosensor using the electrode according to claim 1. 16. The biosensor according to claim 15, wherein the sensor is a glucose sensor. Mixing an enzyme with a self-assembling block copolymer comprising an organic metal and coating the solution on the surface of the electrode,
Here, the self-assembling block copolymer containing the organic metal is self-assembled into an amorphous double continuous structure,
Wherein the crosslinkable block containing a double bond is crosslinked to fix the enzyme. delete delete 18. The method of claim 17, wherein the block copolymer is poly (ferrocenyldimethylsilane-b-isoprene).

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