KR101937883B1 - Binder composition comprising biomaterials - Google Patents

Abstract

A peptide having a binding ability to a carbon material or a binder composition containing a phage displaying the peptide. According to a composition according to an aspect, a carbon material, for example, a peptide or a phage having a strong binding force to a graphitic surface It is possible to enhance the bonding force between the carbon material or the graphitic material or to improve the contact property between the carbon material and the substrate or between the carbon material and the enzyme, thereby realizing a high-performance energy element or a biosensor.

Description

Binder composition comprising biomaterials < RTI ID = 0.0 >

And more particularly to a peptide having a binding ability to a carbon material or a binder composition for forming an electrode comprising a phage on which the peptide is displayed.

Demand for materials and device technologies that can operate flexibly and with high performance, such as wearable computers, flexible displays, flexible batteries, biomedical electrodes and biosensors for health monitoring, and robot-human interfaces are rapidly increasing have. In order to enable such an application, development of a material having excellent mechanical characteristics such as flexibility and excellent electrical characteristics is required. In addition, as in the attachment type biosensor, the development of a material capable of improving the contact property with the active material, such as a material capable of additionally imparting biochemical and biochemical characteristics in addition to the electrical characteristic, or a flexible battery very important. Further, in order to make a high-performance material having these various components combined, a low contact resistance between the components is required, and excellent contact characteristics with a flexible substrate or a flexible current collector are essential. For example, the characteristics of a lithium ion secondary battery are largely influenced by the electrodes, electrolytic solution, and other battery materials used. Among them, interfacial delamination between the current collector and the active material due to external physical stimulation, . Therefore, in a flexible energy element, for example, a flexible secondary battery, it is introduced into the interface between the flexible current collector and the active material, between the active material and the separator, or between the active material particles in the active material layer, There is a need for the development of adhesives that can be used as an adhesive.

Printing processes such as solution-based processes, such as coating processes, screen printing, and inkjet printing, can be mass-produced by a solution process, a variety of materials can be stacked, and the number of processes can be greatly reduced It is widely used in wearable sensor and energy device research due to various advantages such as cost saving because there is no waste of points and materials. Nanocarbon materials such as carbon nanotubes and graphenes have excellent electrical, mechanical and chemical properties. To use these materials with the above-mentioned flexible electronic devices, flexible bioelectrodes, sensors, and flexible energy device electrodes Has recently been actively researched.

Single-walled carbon nanotubes are attracting attention as one of the most suitable materials for fabricating wearable sensor devices due to their unique nano-sized tube structure and their high mechanical, electrical and electrical chemical properties. However, single-walled carbon nanotubes exhibit low dispersion stability in water-based inks and thus show many problems in manufacturing electronic devices through printing. In general, a method of dispersing single-walled carbon nanotubes in water by applying a functional group through a chemical treatment or using a surfactant is mainly used for producing single-walled carbon nanotube ink. When a functional group is applied through a chemical treatment, the electrical and electrochemical characteristics inherent to the carbon nanotubes are deformed by deforming the carbon nanotube structure. Thus, surfactants are mainly used in the fields where the characteristics are important. However, even when a surfactant is used, there are problems that the electrical characteristics of the thin film are impaired due to the presence of a surfactant between the printed carbon nanotubes, and a part of the surfactant is dissolved when coming into contact with water. Wearable biomedical sensors measure the amount of substances present in biological products such as sweat, blood, and tears, and therefore most of them must be in contact with the aqueous solution. Therefore, water stability is an essential factor.

The present inventors have developed a material capable of enhancing bonding between carbon nanotubes and bonding between carbon nanotubes and substrates without deteriorating the electrical properties of carbon nanotubes in order to solve such problems. The M13 phage has the advantage that the peptides on the surface have various types of functional groups and can easily control the types and characteristics of the functional groups. Particularly, a peptide sequence having a strong adhesive force on a graphical surface can be displayed on a phage surface through a bio-panning process or a genetic engineering control. Using this characteristic, bonding between carbon nanotubes and adhesion to a substrate can be improved . The present inventors have completed the invention by developing a carbon nanotube ink using a biomaterial having such adhesive properties.

One aspect is to provide a peptide composition capable of binding to a carbon material, or a binder composition comprising a phage displaying the peptide.

An aspect of the present invention is to provide a composition for electrode printing comprising a peptide capable of binding to a carbon material or a phage displaying the peptide and a carbon material.

Another aspect is to provide a method of printing an electrode using the composition for electrode printing or a method of manufacturing a biosensor.

One aspect provides a binder composition, a composition for electrode printing or a bioadhesive composition, or an adhesive composition for an electrochemical device, which comprises a peptide capable of binding to a carbon material or a phage and a carbon material on which the peptide is displayed.

Another aspect provides an ink composition for ink jet printing comprising a peptide capable of binding to a carbon material or a phage displaying the peptide, and a carbon material.

As used herein, the term " composition for electrode printing " or " ink composition for inkjet printing " is used in an ink-jet printing method in which an electrode is directly patterned by direct printing in the form of a liquid ≪ / RTI > composition. That is, the composition can be mounted on an inkjet printer and printed on a substrate through a nozzle of a printer or the like. Compositions according to one embodiment are distinguished from those laminated by a method such as lithography on a substrate in the form of a sheet or a film. Thus, the composition may be in the form of a liquid. Specifically, the peptide or a solution containing the phage displaying the peptide and a colloidal solution containing the carbon material may be included. In addition, the solution containing the peptide or the phage displaying the peptide may be such that the peptide or the phage is dispersed in the solution. For example, the solution in which the phage is dispersed may contain phage at a concentration of 1 × ^{10 10} / ml to 1 × ^{10 15} / ml. The solution in which the peptide is dispersed may contain a peptide at a concentration of 0.2 mg / ml to 4 mg / ml. In addition, the colloidal solution may contain carbon nanotubes (for example, single-walled carbon nanotubes) at 1 × ^{10 10} / ml to 1 × ^{10 15} / ml. The peptide or the phage-dispersed solution and the colloid solution may be mixed, for example, in a volume ratio of 1: 8 to 8: 1. More specifically, the carbon material and phage may be mixed in a molar ratio of 20: 1 to 1:20.

The composition according to one embodiment is printed on a substrate, for example, inkjet printed and then dried to form an electrode. Further, for example, the electrode may be an electrochemical device, for example, a biosensor device.

The carbon material of the electrode according to one embodiment thus formed is enhanced in the bonding force between the carbon materials or the bonding force between the carbon material and the substrate. Specifically, the peptide or phage promotes the binding between the carbon material or the graphitic material, thereby contributing to the structuring / stabilization of the carbon material or the graphitic material. This is a distinct concept from the functionalization of a graphitic material, which can mean the formation of a network structure of a graphical material. Said carbon material may exhibit increased stability in aqueous solution, for example, as compared to not using the phage on which said peptide or peptide is displayed.

Thus, in one embodiment, the peptide or phage that binds to the graphitic agent may be one that performs a role as a binder composition, or bioadhesive agent. As used herein, " bioadhesive " or " binder composition " may mean that the peptide or phage promotes bonding between carbonaceous material, e.g., a graphical material, or contributes to the adhesion properties of the graphical material and substrate have. Specifically, a phage displaying a peptide capable of binding to a carbon material is specifically bound to a carbon material, and thus can be introduced into an energy element or an interface of each constitution of an electrochemical device to improve the adhesive property or the interface property. For example, when the binder composition is introduced between the current collector and the active material of the energy device, the active material may be further attached to the current collector to thickly coat the active material. Further, for example, when the binder composition is introduced into the substance to be processed of the energy element, the active material can be prevented from peeling at the interface. Accordingly, the binder composition according to one embodiment may improve the adhesion property between the current collector and the active material of the electrochemical device, or between the active material and the separator, or to improve the interface property of the active material. Thus, the binder composition may be used for electrode formation, flexible device, energy device, or electrochemical device. Examples of the energy device include a flexible battery, an alkaline battery, a dry battery, a mercury battery, a lithium battery, a nickel-cadmium battery, a nickel-hydrogen battery, a secondary battery such as a lithium ion secondary battery, And may include a secondary battery.

In another embodiment, the carbon material or the graphitic material is free of charge, and thus a separate functionalization is required in order to attach an enzyme for a biosensor or the like. However, a peptide or phage having a binding ability to a carbon material or a graphical material has a charge The enzyme can be integrated in a network structure of a carbon material or a graphical substance through a coating of an electrolyte or the like. Thus, the phage or peptide may be one that forms a junction of a plurality of carbon materials or a graphitic material. In addition, the peptide may be one in which two peptides are linked by a linker to link two carbon materials or a graphitic substance to each other. In particular, two peptides linked by a linker within the network structure of a plurality of graphitic materials may each be associated with a single graphitic material. The linker may be a peptide linker. The peptide linker may be a variety of linkers known in the art, for example, a linker consisting of a plurality of amino acids. According to one embodiment, the linker may be, for example, a polypeptide consisting of from 1 to 10 or from 2 to 8 any amino acid. The peptide linker may comprise Gly, Asn and Ser residues, and may also include neutral amino acids such as Thr and Ala. Amino acid sequences suitable for peptide linkers are known in the art.

The carbon material may comprise a graphitic material. As used herein, the term " graphitic materials " refers to a material having a graphitic surface on which carbon atoms are arranged in a hexagonal shape. If the material contains a graphitic surface, , Chemical properties, and structural properties of the composition. The graphitic material may be, for example, a graphene sheet, a highly oriented pyrolytic graphite (HOPG) sheet, a single-walled carbon nanotube, or a double-walled carbon nanotube nanotubes, carbon nanotubes such as multi-walled carbon nanotubes, or fullerene. The graphitic material may be a metallic, semiconductive or hybrid material, for example, a mixture of a graphene sheet and a single-walled carbon nanotube.

The peptide can be used without limitation as long as it is a peptide capable of binding to a graphical substance, for example, a carbon substance. For example, X ₂ SX ₁ AAX ₂ X ₃ P (SEQ ID NO: 1), X ₂ X ₂ PX ₃ X ₂ AX ₃ P (SEQ ID NO: 2), SX ₁ AAX ₂ X ₃ P ₂ PX ₃ X ₂ AX ₃ P (SEQ ID NO: 4), or a set of peptides or peptides comprising at least one selected from the group consisting of amino acid sequences of ₂ PX ₃ X ₂ AX ₃ P (SEQ ID NO: 4). The peptide may be one which comprises a conservative substitution of the disclosed peptide. As used herein, the term " conservative substitution " refers to substitution of a first amino acid residue with a second, different amino acid residue, wherein the first and second amino acid residues have a side chain with similar biophysical characteristics It can mean. Similar biophysical properties may include the ability to provide or accept hydrophobic, charge, polar, or hydrogen bonds. Examples of conservative substitutions include, but are not limited to, basic amino acids (arginine, lysine and histidine), acidic amino acids (glutamic and aspartic acid), polar amino acids (glutamine and asparagine), hydrophobic amino acids (leucine, isoleucine, valine and methionine), hydrophilic amino acids Alanine, serine and threonine), aromatic amino acids (phenylalanine, tryptophan, tyrosine and histidine), and small amino acids (glycine, alanine, serine and threonine). In general, amino acid substitutions that do not alter specific activity are known in the art. Thus, for example, in the peptide X ₁ can be W, Y, F or H, X ₂ can be D, E, N or Q, and X ₃ can be I, L or V. In addition, the peptide may be a peptide or a peptide set comprising at least one selected from the group consisting of the amino acid sequences of SEQ ID NOS: 5 to 11. Peptides other than phage-derived, for example, peptides comprising the amino acid sequence of SEQ ID NO: 12 may also be used. The peptide may be one in which two peptides are linked through a linker (e. G., SEQ ID NO: 10 or SEQ ID NO: 11). The peptide may also further comprise a portion of the epitope protein of the phage, for example, 1 to 10 amino acid residues (e. G., SEQ ID NO: 11). The N-terminal or C-terminal end of the amino acid sequence of the peptide or peptide set may be linked to the consecutive amino acid sequence of the phage coat protein. Thus, for example, the set of peptides or peptides may comprise a sequence of 5 to 60 amino acids, 7 to 55 amino acids, 7 to 40 amino acids, 7 to 30 amino acids, 7 to 20 amino acids, 7 to 10 amino acid sequences. The peptide or set of peptides may be an assembled (e. G., Self-assembled) peptide or set of peptides. For example, the peptide or peptide set may be composed of a structure of? -Helix or? -Sheet. The peptide or peptide set can improve the binding between the graphical materials so that the graphical material has a mesh structure.

Peptides that bind to the graphitic agent can be selected through a library of peptides and can be selected, for example, through phage display techniques. The phage display technique allows the peptide to be displayed on the outside of the phage genetically linked, inserted, or substituted into the phage's coat protein, and the peptide can be encoded by the genetic information in the virion. By screening the proteins of various variants by the displayed proteins and the DNA encoding them, they can be screened and called "biopanning". Briefly, the bio-panning technique involves reacting a displayed phage with an immobilized target (e.g., a graphical material), washing the unbound phage, and then destroying the binding interaction between the phage and the target, And a method of eluting the combined phage. A portion of the eluted phage can be left for DNA sequencing and peptide identification, and the remainder can be amplified in vivo and a sub-library for the next round can be generated and repeated.

The peptide may also be displayed on the envelope protein of the phage. Thus, for example, a phage that is capable of binding to the glycocalic material may be one comprising the peptidic protein of phage or the peptide displayed on the fragment.

The term " phage " or " bacteriophage " is used interchangeably and can refer to a virus that infects bacteria and replicates in bacteria. Phage or bacteriophage may be used to display peptides that selectively or specifically bind to a graphical substance or a volatile organic compound. The phage may be one that has been genetically engineered such that a peptide capable of binding to the graphical substance is displayed on the envelope protein or fragment thereof of the phage. The term " genetic engineering " or " genetically engineered " in the context of the present invention is intended to include peptides having a binding capacity to a glycotic material, May refer to the act of introducing a genetic modification or a phage created thereby. The genetic modification includes introducing a foreign gene encoding the peptide. The phage may be a filamentous phage, for example, M13 phage, F1 phage, Fd phage, If1 phage, Ike phage, Zj / Z phage, Ff phage, Xf phage, Pf1 phage, or Pf3 It can be a phage.

The term " phage display " in the present invention may refer to the display of functional foreign peptides or proteins on the surface of phage or phagemid particles. The surface of the phage may mean the envelope protein of the phage or a fragment thereof. Also, the phage may be linked to the N-terminus of the phage coat protein of the phage, or the C-terminus of the functional peptide may be inserted between consecutive amino acid sequences of the phage coat protein of the phage or the consecutive amino acid sequence of the coat protein Which is a part of the phage. The position of the consecutive amino acid sequence in which the peptide is inserted or substituted in the coat protein is selected from the N-terminal of the coat protein at positions 1 to 50, positions 1 to 40, positions 1 to 30, positions 1 to 20, Position 10, position 2 to 8, position 2 to 4, position 2 to 3, position 3 to 4, or position 2. In addition, the envelope protein may be p3, p6, p8 or p9. For example, the C-terminal of any one of SEQ ID NOS: 1 to 12 may be linked to the N-terminal of p8 (SEQ ID NO: 18) having a length of 50 amino acids present in the trunk of the M13 phage. For example, when the peptide of any one of SEQ ID NOS: 1 to 12 is an amino acid sequence (i.e., EGD) at position 2 to 4 of the coat protein p8 of M13 phage, position 2 to 3, position 3 to 4 Position, or in place of the amino acid sequence of position 2.

In other embodiments, the phage can be directionally arranged on the surface of the carbonaceous material using the structure of the filamentary form of the phage itself. For example, they may be aligned in a specific direction, in which case the binding force between the peptide located on the coat protein of the phage and the surface of the carbonaceous material may increase and be aligned on the same day. A phage aligned in a day can impart anisotropic functionalization to the surface of the carbonaceous material, which is different from isotropic or random functionalization only when the peptide alone is used. In addition to the above ordered alignment structure, a structure having a specific directionality such as a smectic structure, a layered structure such as a nematic structure, a spiral structure, and a lattice structure can be formed, Various functions can be given on the surface.

In another embodiment, the graphitic material may comprise a network structure, and the enzyme may be on the network structure, within the network structure, and / or below the network structure. In addition, the network structure may be a binding complex of the graphitic substance and the peptide or a binding complex of the graphitic substance and the phage. Accordingly, the internal structure of the graphitic material may have a percolated network structure. As used herein, the term " percolated network " may refer to a lattice structure composed of randomly conducting or non-conducting connections.

In another aspect, the method includes printing the composition for electrode printing on a substrate, for example, inkjet printing; And drying the printed composition to form an electrode.

In another aspect, the method includes printing the composition for electrode printing on a substrate, for example, inkjet printing; Drying the printed composition to form an electrode; And forming an enzyme on the formed electrode. The present invention also provides a method for manufacturing a biosensor.

The composition for electrode printing or the ink composition for inkjet printing is as described above.

The enzyme may be an analyte binding material. The term " analyte binding materials " or " analyte binding reagents " in the context of the present invention may be used interchangeably and refer to an analyte-specifically binding substance. The binding substance may include a redox enzyme. The redox enzyme may mean an enzyme that oxidizes or reduces a substrate, and may be, for example, an oxidase, peroxidase, reductase, catalase, or di Examples of the oxidoreductase include glucose oxidase, lactate oxidase, cholesterol oxidase, glutamate oxidase, horseradish peroxidase (HRP), alcohol oxidase, glucose oxidase (GOx), glucose dihydrogenase Glucose dehydrogenase (GDH), cholesterol ester synthase, ascorbic acid oxidase ( ascorbic acid oxidase, alcohol dehydrogenase, laccase, tyrosinase, galactose oxidase or bilirubin oxidase. The enzyme may be a glycocalypse substance The term " immobilized " may mean a chemical or physical association between the enzyme and the graphitic substance. The term " immobilized "

The method of printing the electrode or the method of manufacturing the biosensor may further include processing the polymer material on the electrode. As described above, the composition according to an exemplary embodiment may not require a separate modification, but may be modified with a positive charge polymer or a negative charge polymer so as to additionally have a positive charge or a negative charge. Examples of the positive charge polymer include PAH (poly (allyamine), PDDA (poly (dimethyliminammonium)), PEI (polyimide), or PAMPDDA (acrylamide-co-diallyldimethylammonium) Examples of which are PSS (poly (4-styrenesulfonate), PAA (poly (acrylic acid), PAM (poly (acryl amide), poly (vinylphosphonic acid) propanesulfonic acid), PATS (poly (anetholesulfonic acid)), or PVS (poly (vinyl sulfate)).

The substrate may be a conductive substrate or an insulating substrate. The electrode may be a first electrode, a second electrode, or a third electrode, and may be a working electrode, a counter electrode, or a reference electrode. In addition to the working electrode, the counter electrode, or the reference electrode, the electrode may further include an auxiliary electrode and a recognition electrode.

According to one aspect of the present invention, by using a carbon material, for example, a peptide or a phage having a strong binding force to a graphitic surface, the binding force between the carbon material or the graphitic material is enhanced, Or the contact property between the carbon material and the enzyme is improved, thereby realizing a high-performance energy element or a biosensor.

FIG. 1 is an image showing the difference in stability in an aqueous solution of a film produced using the bioadhesive (Production Examples 1 to 4) according to one embodiment and a film produced using the solution of Comparative Example 1. FIG.
2 is an image of a spray of a bioadhesive according to one embodiment.
FIG. 3 is a graph showing a change in sheet resistance with time in an electrode prepared by printing a bioadhesive according to one embodiment (Production Example 1) and an electrode produced by printing a solution of Comparative Example 1 in distilled water. FIG.
4 is a view showing an electrode for a biosensor printed using a bio-adhesive according to one embodiment.
5 is a view showing an electrode for a multi-biosensor printed using a bio-adhesive according to one embodiment.
Figure 6 is a graph showing the direct electron transfer peak of a glucose sensor according to one embodiment.
7 is a graph showing a redox curve according to a scanning speed of a glucose sensor according to an embodiment.
8 is a graph showing a redox peak current value according to a scanning speed of a glucose sensor according to an embodiment.
9 is a graph showing a redox curve according to glucose concentration of an all-printed glucose sensor according to one embodiment.
10 is a graph showing a change in peak of reduction current according to glucose concentration in an all-printed glucose sensor according to one embodiment.
11 is a graph showing the detection performance of glucose in human serum or artificial tears of a glucose sensor according to one embodiment.
12 is a graph showing the redox curve according to the lactate concentration of the lactate sensor according to one embodiment.
13 is a graph showing a change in the peak reduction current value according to the lactate concentration of the lactate sensor according to one embodiment.
FIG. 14 is a graph showing changes in current due to the addition of glucose and lactate in a multi-biosensor according to an embodiment.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

 [ Manufacturing example  1] Preparation of bio-adhesive containing phage

As one embodiment of the present invention, a bioadhesive solution was prepared by the following method.

Preparation of phage solution

(GP2) displaying the M13 phage (GP1) and the NPIQAVP (SEQ ID NO: 6) displaying the SWAADIP (SEQ ID NO: 5), which is a peptide having a strong binding force with the M13 phage, Was prepared by the following method.

First, an M13HK vector was constructed by site-directed mutation of C131 base pair (C13) of the M13KE vector (NEB, product # N0316S, SEQ ID NO: 13)

Herein, the M13KE vector (NEB, product # N0316S) is a cloning vector (cloning vector M13KE) consisting of 7222 bp DNA, and the genetic information is available on the Internet site (https://www.neb.com/~/media/NebUs/Page% 20Images / Tools% 20and% 20Resources / Interactive% 20Tools / DNA% 20Sequences% 20and% 20Maps / Text% 20Documents / m13kegbk.txt). The nucleotide sequences of the oligonucleotides used in the site-specific mutation are as follows:

5'-AAG GCC GCT TTT GCG GGA TCC TCA CCC TCA GCA GCG AAA GA-3 '(SEQ ID NO: 14), and

5'-TCT TTC GCT GCT GAG GGT GAG GAT CCC GCA AAA GCG GCC TT-3 '(SEQ ID NO: 15).

A phage display p8 peptide library was constructed using the restriction enzyme BspHI (NEB, product # R0517S) and BamHI restriction enzyme (NEB, product # R3136T) in the prepared M13HK vector.

The nucleotide sequence of the oligonucleotides used in the preparation of the phage display p8 peptide library as an embodiment of the present invention is as follows:

5'-TTA ATG GAA ACT TCC TCA TGA AAA AGT CTT TAG TCC TCA AAG CCT CTG TAG CCG TTG CTA CCC TCG TTC CGA TGC TGT CTT TCG CTG CTG -3 '(SEQ ID NO: 16), and

5'-AAG GCC GCT TTT GCG GGA TCC NNM NNM NNM NNM NNM NNM NCA GCA GCG AAA GAC AGC ATC GGA ACG AGG GTA GCA ACG GCT ACA GAG GCT TT -3 '(SEQ ID NO: 17).

The nucleotide sequence of the prepared phage display p8 peptide library was 4.8 × 10 ⁷ pfu (plaque form unit) and had a copy number of about 1.3 × 10 ⁵ per each sequence.

Then, the prepared phage display p8 peptide library was bound to a graphical surface by a bio-panning method to select a phage displaying a peptide to be used as the biomaterial of the present invention. The bio-panning method is specifically as follows.

First, HOPG (highly oriented pyrolytic graphite, SPI, product # 439HP-AB), a material with a graphical surface, was peeled off with tape before the experiment to obtain a fresh surface to minimize defects such as oxidation of the sample surface. At this time, the HOPG substrate used was a relatively large HOPG substrate having a grain size of 100 μm or less.

The prepared phage display p8 peptide library of 10.8 × 10 ¹⁰ (4.8 × 10 ⁷ replicates, 1000 copies per sequence) phage display library was prepared in 100 μl of Tris-Buffered Saline buffer, Lt; / RTI > in a shaking incubator at < RTI ID = 0.0 > 100 < / RTI > After 1 hour, the solution was removed and then washed 10 times in TBS. The washed HOPG surface was reacted with Tris-HCl (pH 2.2) as an acid buffer for 8 minutes to elute the weakly reactive peptide, and then subjected to a mid-log XL-1 blue E. coli culture The culture was eluted for 30 minutes. A portion of the eluted culture was left for DNA sequencing and peptide identification and the rest amplified to create a sub-library for the next round. The above procedure was repeated using the created sub-library. On the other hand, the plaque left was analyzed for DNA to obtain a p8 peptide sequence. Analysis of the obtained sequence revealed that the phage (GP1) and the NPIQAVP (SEQ ID NO: 5) displaying the peptide sequence SWAADIP (SEQ ID NO: 6) was displayed.

Preparation of colloidal solution

First, an aqueous solution prepared by adding sodium or cholate as a surfactant to the distilled water at a concentration of 1 or 2% w / v was prepared. Then, single-walled carbon nanotubes (manufactured by Nanointegris, SuperPure SWNTs, solution Type, concentration: 250 μg / ml or 1000 μg / ml) was dialyzed for 48 hours to stabilize the single-walled carbon nanotubes with sodium cholate to prepare a colloidal solution.

Assuming that the average length of carbon nanotubes (CNTs) is 1 μm and the average diameter is 1.4 nm, the calculation formula of the number of single-walled carbon nanotubes contained in the colloid solution is as follows.

[Equation 1]

Number of single-walled carbon nanotubes (pcs / ml) = concentration μg / ml X 3 x 10 ¹¹ CNT

According to the above equation, it can be seen that the number of single-walled carbon nanotubes contained in the 1000 μg / ml colloid solution is 3 × ^{10 14} / ml. The number of single-walled carbon nanotubes per volume was controlled using an aqueous solution of sodium cholate at the same concentration as the dialyzed solution.

Manufacture of bio-adhesives

TBS (Tris-Buffered saline) the M13 phage (GP1) having a surface and a strong bonding force of the material in our tick buffer was dispersed at a concentration of 6X10 ¹³ gae / ml. The colloid solution prepared in Preparation Example 1 and the M13 phage solution were mixed at a volume ratio of 2: 1 to mix the graphite material and M13 phage (GP1) in a molar ratio of 10: 1. At this time, the number of M13 grains contained in the M13 phage solution was calculated by the following empirical formula. A _269nm and _{A320 nm} represents the absorbance of the solution at the wavelength of the light, 269 nm and 320 nm.

&Quot; (2) "

[ Manufacturing example  2-4] Preparation of bioadhesive using phage-derived peptide

A phage-derived peptide that specifically binds to the graphitic surface was chemically synthesized (manufactured by Peptron, Inc.) at a concentration of 1 mg / mL. Specifically, three types of phage-derived peptides were used. (SEQ ID NO: 9) (ADSWAADIPDPA) (Preparation Example 2), peptide sequence No. 10 (ADSWAADIPDPAGGG ADSWAADIPDPA) (Preparation Example 3) linking two peptides SEQ ID NO: 9 with a linker, and a partial sequence (KAA) A peptide sequence No. 11 (ADSWAADIPDPAKAAGGGADSWAADIPDPAKAA) (Preparation Example 4) in which two peptides were linked by a linker was used. Two peptide sequences were linked by a linker to allow each peptide to bind to one graphical substance and consequently to connect two graphical substances. 1 mL of the prepared single-stranded carbon nanotube colloid solution (3.0 X 10 ¹⁴ / mL) prepared by the method described in Preparation Example 1 was mixed with 1 mL of the peptide of SEQ ID NOs: 9 to 11 prepared above.

[ Comparative Example  1] Preparation of ink solution containing only colloidal solution

The colloid solution prepared in Preparation Example 1 was used.

[ Experimental Example  1] Film made with bio-adhesive Comparative Example  Evaluation of the stability of a film in an aqueous solution using a solution of 1

(Single-walled carbon nanotubes 1000 μg / ml, sodium cholate 1 w / v%) prepared in Production Examples 1 to 4 and the colloid solution prepared in Comparative Example 1 (single-walled carbon nanotubes 1000 μg / ml, sodium 5 μL of a commercially available polyethylene terephthalate (PET) film was dried by using an ink containing only 1 w / v% of cholate and then put into distilled water to confirm the change after 6 minutes. Thus, The results are shown in Fig.

As shown in FIG. 1, the films prepared using the bioadhesives prepared in Production Examples 1 to 4 were confirmed to maintain their original shape in distilled water. On the other hand, the films prepared using the colloid prepared in Comparative Example 1, And it can be confirmed that a lot of existing shapes are collapsed. As a result, it can be seen that the single-walled carbon nanotube film produced using the bio-adhesive exhibits high stability in an aqueous solution.

[ Experimental Example  2] Sensor electrode manufactured using bio-adhesive Comparative Example  Stability Evaluation of Electrical Conductivity of Electrode Using Solution of 1

(Single-stratified carbon nanotube 250 μg / ml, sodium cholate 2 w / v%) prepared in Preparation Example 1 and the colloid solution (single stratum carbon nanotube 250 μg / ml, sodium cholate 2 (Hansol) photo paper (210 g / m2) using inks containing only a small amount of ink (5% w / v%). The injection image of the ink containing the bio-adhesive is shown in Fig. Then, the electrode pattern was placed in distilled water at predetermined time intervals, and the sheet resistance was measured to evaluate the stability of the electrode's electrical conductivity in aqueous solution. The sheet resistance was measured by the Van der Pauw method, and the results are shown in FIG.

As shown in FIG. 3, in the case of the electrode manufactured using the bioadhesive, the sheet resistance was increased by 35% when compared with that in the distilled water when the electrode was put out in distilled water for 20 minutes, Did not do it. On the other hand, in the case of the electrode prepared using the ink containing only the colloid solution, the sheet resistance increased by 293% in the 5th printing and 143% in the 10th printing. As a result, it can be seen that the sensor electrode made of the bioadhesive has a high conductivity and a high electrical conductivity stability in an aqueous solution.

[ Manufacturing example  5] Using bio-adhesive Third generation Glucose  Manufacture of sensors

The bioadhesive prepared in Preparation Example 1 was printed on a photo paper glossy 210 μm (Hansol) commercially available through ink jet printing. The shape of the printed biosensor is shown in Fig. As shown in FIG. 4, the electrode for a biosensor includes a working electrode, a reference electrode, and a counter electrode.

After printing the electrodes, 5 μL of a 5 w / v% aqueous polyethyleneimine (PEI) solution was dropped onto the working electrode of the printed electrode and dried. After drying was completed, the excess PEI was washed away using distilled water. Then, 5 μL of an aqueous solution of glucose oxidase enzyme (GOx) at a concentration of 100 mg / ml was further dropped on a working electrode and dried to prepare a third generation glucose sensor.

[ Manufacturing example  6] Using bio-adhesive Third generation Lactate  Manufacture of sensors

An electrode for a biosensor was printed in the same manner as in Production Example 5. Then 5 μL of a 5 w / v% polyethyleneimine (PEI) aqueous solution was dropped onto the working electrode of the printed electrode and dried. After drying was completed, the excess PEI was washed away using distilled water. 5 μL of an aqueous solution of lactate oxidase enzyme (LOx) at a concentration of 100 mg / ml was further dropped on a working electrode and dried to produce a third generation lactate sensor.

[ Manufacturing example  7] All-printed Third generation Glucose  Manufacture of sensors

An electrode was printed in the shape of FIG. 4 using a bioadhesive in the same manner as in Production Example 5 above. Then, 5 w / v% solution of PEI was printed on the working electrode through the inkjet printing method and dried. After drying was completed, the excess PEI was washed away using distilled water. An all-printed third-generation glucose sensor was prepared by printing a GOx enzyme aqueous solution at a concentration of 100 mg / ml on the working electrode 10 times.

[ Manufacturing example  8] Manufacture of multi-biosensor using bio-adhesive

The ink containing the bioadhesive prepared in Preparation Example 1 was printed on a photo paper glossy 210 .mu.m (Hansol), which can be commercially purchased through an inkjet printing method. The shape of the printed multi-biosensor is shown in Fig. As shown in FIG. 5, an electrode for a multi-biosensor is composed of two working electrodes and a counter electrode.

After printing electrodes for multiple biosensors, 5 w / v% PEI solution was dropped onto the two working electrodes and dried. After drying was completed, the excess PEI was washed away using distilled water. 5 μL of a GOx enzyme aqueous solution at a concentration of 100 mg / ml and 5 μL of an aqueous LOx enzyme solution at a concentration of 100 mg / mL were dropped on each of the electrodes shown in FIG. 5 and dried to produce a multi-biosensor capable of measuring glucose and lactate Respectively.

[ Experimental Example  3] Measurement of electrochemical activity of sensor electrode fabricated with bio-adhesive

The glucose sensor prepared in Preparation Example 5 was injected with a negative voltage of -0.6 V to -0.1 V at a rate of 200 mV / s in a 10 mM PBS buffer (pH = 7.4, 79383, Sigma Aldrich) The results are shown in Fig.

As shown in FIG. 6, the produced glucose sensor shows a strong redox peak on the cyclic voltammetry (CV) in the range of about -400 mV as compared to the Ag / AgCl reference electrode (3 M KCl saturated, PAR, K0260) have. This implies that the direct-electron-transfer (DET) reaction takes place as shown in the following equation since the FAD redox center in GOx forms an efficient / direct electrical pair with single-walled carbon nanotubes .

FAD + 2H ⁺ + 2e ^- -> FADH ₂

The results are shown in FIG. 7, and the peak value of the redox current with respect to each scanning speed is shown in FIG. 8 .

As shown in FIG. 8, when the voltage scanning speed was increased from 20 mV / s to 2000 mV / s in a sequential manner, the redox current was linearly increased, confirming the surface control reaction. Based on this reaction, it can be seen that the enzyme present in the biosensor electrode manufactured using the bioadhesive can efficiently transfer electrons directly to the electrode.

[ Experimental Example  4] manufactured by using bio-adhesive GOx  Of the enzyme electrode To glucose  Evaluation of Reactivity

CV was measured while the glucose sensor prepared in Preparation Example 5 was injected with a voltage of 200 mV / s on a 10 mM PBS buffer (pH = 7.4, 79383, Sigma Aldrich) solution containing 10 μM to 7500 μM of glucose , And the results are shown in Fig.

As shown in FIG. 10, when the glucose concentration contained in the 10 mM PBS buffer was increased in the state where the voltage was applied between -0.1 V and -0.6 V, the reduction current was linearly increased to the glucose concentration of 1000 uM As shown in Fig. The sensitivity of this measured glucose sensor was measured to be ~ 62.1 μA / mM cm ² . At glucose concentrations higher than 2500uM, there was little change in the reduction electrode, which is a result of the higher concentration of glucose in solution than the glucose concentration that the enzyme contained in the electrode can handle, increasing the amount of enzyme contained in the electrode The amount of glucose that can be analyzed can be further increased. As a result, the DET-based glucose sensor according to the present invention exhibits a reduction current (100 μM to 500 μM) in a glucose concentration range (100 μM to 500 μM) contained in body fluids (sweat, tears, needles, etc.) Can be used as a DET-based third-generation wearable biosensor.

[ Experimental Example  5] manufactured by using bio-adhesive GOx  Of the enzyme electrode To glucose  Selectivity for

The selectivity of the glucose sensor prepared in Preparation Example 5 for the glucose reaction was evaluated. Specifically, for 10 mM glucose in 10 mM PBS, 10 mM glucose in PBS, human serum (~11 mM) in PBS, and 10 mM glucose in artificial tears, a negative voltage of -0.6 V to -0.1 V at 200 mV / s And the results are shown in FIG. 11. The results are shown in FIG. Since artificial tears and human serum contain glucose as well as many other inhibitory factors, the selectivity of the glucose sensor can be assessed by comparing the response of the glucose sensor to these solutions.

As shown in FIG. 11, the third-generation biosensor of the GOx enzyme electrode manufactured using the bioadhesive according to one embodiment is stable in a physiological environment such as human serum or artificial tears as well as a 10 mM PBS solution as a buffer solution , It can be seen that it is possible to drive in the solution of the non-invasive method such as needle / tear / sweat.

[ Experimental Example  6] manufactured using bio-adhesive LOx  Of the enzyme electrode On lactate  Evaluation of Reactivity

The lactate sensor prepared in Preparation Example 6 was injected at a rate of 200 mV / s on a solution of 10 mM PBS buffer (pH = 7.4, 79383, Sigma Aldrich) containing 2500 μM of lactate at 10 μM, And the results are shown in Figs. 12 and 13. Fig.

As shown in FIG. 12, a strong redox peak appears on the cyclic voltammetry (CV) at about -375 mV compared to the Ag / AgCl reference electrode. The above results indicate that the FAD redox center in the LOx can be efficiently transferred directly to the single-walled carbon nanotubes in the printing electrode (direct-electron-transfer, DET). In addition, when the lactate concentration contained in the 10 mM PBS buffer was increased while the voltage was applied between -0.2 V and -0.6 V, as shown in FIG. 13, the reduction current linearly increased in the positive direction It can be seen that The sensitivity of the measured lactate sensor was measured to be ~ 116 μA / mM cm ² .

[ Experimental Example  7] Characteristic evaluation of multi-biosensor fabricated using bio-adhesive

-0.4 V was applied to the working electrode of the GOx, LOx enzyme-based multi-biosensor manufactured in Production Example 8, and the current from each electrode was measured. The results are shown in FIG.

As shown in Fig. 14, an increase in current in the positive direction was observed at the GOx enzyme working electrode when glucose was added, whereas no increase in current was observed at the LOx enzyme working electrode. In addition, when lactate was added, an increase in current was observed at the LOx enzyme working electrode, but no increase in current was observed at the GOx enzyme working electrode. At the same time, when glucose and lactate were added, an increase in current was observed at both working electrodes at the same time. This means that the GOx enzyme working electrode and the LOx enzyme working electrode are induced to increase the current due to the enzyme reaction only on each substrate. Therefore, the sensor according to one embodiment is not only highly selectable, It can be seen that it can also be used as a sensor.

<110> Korea Institute of Science and Technology <120> Binder composition comprising biomaterials <130> PN115443 <160> 18 <170> PatentIn version 3.5 <210> 1 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> peptide selectively binding to graphitic materials <220> <221> VARIANT <222> (1) <223> X is D, E, N, or Q <220> <221> VARIANT <222> (3) <223> X is W, Y, F or H <220> <221> VARIANT <222> (6) <223> X is D, E, N or Q <220> <221> VARIANT <222> (7) <223> X is I, L, or V <400> 1 Xaa Ser Xaa Ala Ala Xaa Xaa Pro   1 5 <210> 2 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> peptide selectively binding to graphitic materials <220> <221> VARIANT <222> (1) <223> X is D, E, N, or Q <220> <221> VARIANT <222> (2) <223> X is D, E, N, or Q <220> <221> VARIANT <222> (4) <223> X is I, L, or V <220> <221> VARIANT <222> (5) <223> X is D, E, N, or Q <220> <221> VARIANT <222> (7) <223> X is I, L, or V <400> 2 Xaa Xaa Pro Xaa Xaa Ala Xaa Pro   1 5 <210> 3 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> peptide selectively binding to graphitic materials <220> <221> VARIANT <222> (2) <223> X is W, Y, F, or H <220> <221> VARIANT <222> (5) <223> X is D, E, N, or Q <220> <221> VARIANT <222> (6) <223> X is I, L, or V <400> 3 Ser Xaa Ala Ala Xaa Xaa Pro   1 5 <210> 4 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> peptide selectively binding to graphitic materials <220> <221> VARIANT <222> (1) <223> X is D, E, N, or Q <220> <221> VARIANT <222> (3) <223> X is I, L, or V <220> <221> VARIANT <222> (4) <223> X is D, E, N, or Q <220> <221> VARIANT <222> (6) <223> X is I, L, or V <400> 4 Xaa Pro Xaa Xaa Ala Xaa Pro   1 5 <210> 5 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> peptide selectively binding to graphitic materials <400> 5 Asp Ser Trp Ala Ala Asp Ile Pro   1 5 <210> 6 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> peptide selectively binding to graphitic materials <400> 6 Asp Asn Pro Ile Gln Ala Val Pro   1 5 <210> 7 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> peptide selectively binding to graphitic materials <400> 7 Ser Trp Ala Ala Asp Ile Pro   1 5 <210> 8 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> peptide selectively binding to graphitic materials <400> 8 Asn Pro Ile Gln Ala Val Pro   1 5 <210> 9 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> peptide selectively binding to graphitic materials <400> 9 Ala Asp Ser Trp Ala Ala Asp Asp Pro Asp Pro Ala   1 5 10 <210> 10 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> peptide selectively binding to graphitic material <400> 10 Ala Asp Ser Trp Ala Ala Asp Ile Pro Asp Pro Ala Gly Gly Gly Ala   1 5 10 15 Asp Ser Trp Ala Ala Asp Ile Pro Asp Pro Ala              20 25 <210> 11 <211> 33 <212> PRT <213> Artificial Sequence <220> <223> peptide selectively binding to graphitic material <400> 11 Ala Asp Ser Trp Ala Ala Asp Ile Pro Asp Pro Ala Lys Ala Ala Gly   1 5 10 15 Gly Gly Ala Asp Ser Trp Ala Ala Asp Ile Pro Asp Pro Ala Lys Ala              20 25 30 Ala     <210> 12 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> peptide selectively binding to graphitic materials <400> 12 Tyr Tyr Ala Cys Ala Tyr Tyr   1 5 <210> 13 <211> 7222 <212> DNA <213> Artificial Sequence <220> <223> cloning vector M13KE <400> 13 aatgctacta ctattagtag aattgatgcc accttttcag ctcgcgcccc aaatgaaaat 60 atagctaaac aggttattga ccatttgcga aatgtatcta atggtcaaac taaatctact 120 cgttcgcaga attgggaatc aactgttata tggaatgaaa cttccagaca ccgtacttta 180 gttgcatatt taaaacatgt tgagctacag cattatattc agcaattaag ctctaagcca 240 tccgcaaaaa tgacctctta tcaaaaggag caattaaagg tactctctaa tcctgacctg 300 ttggagtttg cttccggtct ggttcgcttt gaagctcgaa ttaaaacgcg atatttgaag 360 tctttcgggc ttcctcttaa tctttttgat gcaatccgct ttgcttctga ctataatagt 420 cagggtaaag acctgatttt tgatttatgg tcattctcgt tttctgaact gtttaaagca 480 tttgaggggg attcaatgaa tatttatgac gattccgcag tattggacgc tatccagtct 540 aaacatttta ctattacccc ctctggcaaa acttcttttg caaaagcctc tcgctatttt 600 gt; aattcctttt ggcgttatgt atctgcatta gttgaatgtg gtattcctaa atctcaactg 720 atgaatcttt ctacctgtaa taatgttgtt ccgttagttc gttttattaa cgtagatttt 780 tcttcccaac gtcctgactg gtataatgag ccagttctta aaatcgcata aggtaattca 840 caatgattaa agttgaaatt aaaccatctc aagcccaatt tactactcgt tctggtgttt 900 ctcgtcaggg caagccttat tcactgaatg agcagctttg ttacgttgat ttgggtaatg 960 aatatccggt tcttgtcaag attactcttg atgaaggtca gccagcctat gcgcctggtc 1020 tgtacaccgt tcatctgtcc tctttcaaag ttggtcagtt cggttccctt atgattgacc 1080 gtctgcgcct cgttccggct aagtaacatg gagcaggtcg cggatttcga cacaatttat 1140 caggcgatga tacaaatctc cgttgtactt tgtttcgcgc ttggtataat cgctgggggt 1200 caaagatgag tgttttagtg tattcttttg cctctttcgt tttaggttgg tgccttcgta 1260 gtggcattac gtattttacc cgtttaatgg aaacttcctc atgaaaaagt ctttagtcct 1320 caaagcctct gtagccgttg ctaccctcgt tccgatgctg tctttcgctg ctgagggtga 1380 cgatcccgca aaagcggcct ttaactccct gcaagcctca gcgaccgaat atatcggtta 1440 tgcgtgggcg atggttgttg tcattgtcgg cgcaactatc ggtatcaagc tgtttaagaa 1500 attcacctcg aaagcaagct gataaaccga tacaattaaa ggctcctttt ggagcctttt 1560 ttttggagat tttcaacgtg aaaaaattat tattcgcaat tcctttagtg gtacctttct 1620 attctcactc ggccgaaact gttgaaagtt gtttagcaaa atcccataca gaaaattcat 1680 ttactaacgt ctggaaagac gacaaaactt tagatcgtta cgctaactat gagggctgtc 1740 tgtggaatgc tacaggcgtt gtagtttgta ctggtgacga aactcagtgt tacggtacat 1800 gggttcctat tgggcttgct atccctgaaa atgagggtgg tggctctgag ggtggcggtt 1860 ctgagggtgg cggttctgag ggtggcggta ctaaacctcc tgagtacggt gatacaccta 1920 ttccgggcta tacttatatc aaccctctcg acggcactta tccgcctggt actgagcaaa 1980 accccgctaa tcctaatcct tctcttgagg agtctcagcc tcttaatact ttcatgtttc 2040 agaataatag gttccgaaat aggcaggggg cattaactgt ttatacgggc actgttactc 2100 aaggcactga ccccgttaaa acttattacc agtacactcc tgtatcatca aaagccatgt 2160 atgacgctta ctggaacggt aaattcagag actgcgcttt ccattctggc tttaatgagg 2220 atttatttgt ttgtgaatat caaggccaat cgtctgacct gcctcaacct cctgtcaatg 2280 ctggcggcgg ctctggtggt ggttctggtg gcggctctga gggtggtggc tctgagggtg 2340 gcggttctga gggtggcggc tctgagggag gcggttccgg tggtggctct ggttccggtg 2400 attttgatta tgaaaagatg gcaaacgcta ataagggggc tatgaccgaa aatgccgatg 2460 aaaacgcgct acagtctgac gctaaaggca aacttgattc tgtcgctact gattacggtg 2520 ctgctatcga tggtttcatt ggtgacgttt ccggccttgc taatggtaat ggtgctactg 2580 gtgattttgc tggctctaat tcccaaatgg ctcaagtcgg tgacggtgat aattcacctt 2640 taatgaataa tttccgtcaa tatttacctt ccctccctca atcggttgaa tgtcgccctt 2700 ttgtctttgg cgctggtaaa ccatatgaat tttctattga ttgtgacaaa ataaacttat 2760 tccgtggtgt ctttgcgttt cttttatatg ttgccacctt tatgtatgta ttttctacgt 2820 ttgctaacat actgcgtaat aaggagtctt aatcatgcca gttcttttgg gtattccgtt 2880 attattgcgt ttcctcggtt tccttctggt aactttgttc ggctatctgc ttacttttct 2940 taaaaagggc ttcggtaaga tagctattgc tatttcattg tttcttgctc ttattattgg 3000 gcttaactca attcttgtgg gttatctctc tgatattagc gctcaattac cctctgactt 3060 tgttcagggt gttcagttaa ttctcccgtc taatgcgctt ccctgttttt atgttattct 3120 ctctgtaaag gctgctattt tcatttttga cgttaaacaa aaaatcgttt cttatttgga 3180 ttgggataaa taatatggct gtttattttg taactggcaa attaggctct ggaaagacgc 3240 tcgttagcgt tggtaagatt caggataaaa ttgtagctgg gtgcaaaata gcaactaatc 3300 ttgatttaag gcttcaaaac ctcccgcaag tcgggaggtt cgctaaaacg cctcgcgttc 3360 ttagaatacc ggataagcct tctatatctg atttgcttgc tattgggcgc ggtaatgatt 3420 cctacgatga aaataaaaac ggcttgcttg ttctcgatga gtgcggtact tggtttaata 3480 cccgttcttg gaatgataag gaaagacagc cgattattga ttggtttcta catgctcgta 3540 aattaggatg ggatattatt tttcttgttc aggacttatc tattgttgat aaacaggcgc 3600 gttctgcatt agctgaacat gttgtttatt gtcgtcgtct ggacagaatt actttacctt 3660 ttgtcggtac tttatattct cttattactg gctcgaaaat gcctctgcct aaattacatg 3720 ttggcgttgt taaatatggc gattctcaat taagccctac tgttgagcgt tggctttata 3780 ctggtaagaa tttgtataac gcatatgata ctaaacaggc tttttctagt aattatgatt 3840 ccggtgttta ttcttattta acgccttatt tatcacacgg tcggtatttc aaaccattaa 3900 atttaggtca gaagatgaaa ttaactaaaa tatatttgaa aaagttttct cgcgttcttt 3960 gtcttgcgat tggatttgca tcagcattta catatagtta tataacccaa cctaagccgg 4020 aggttaaaaa ggtagtctct cagacctatg attttgataa attcactatt gactcttctc 4080 agcgtcttaa tctaagctat cgctatgttt tcaaggattc taagggaaaa ttaattaata 4140 gcgacgattt acagaagcaa ggttattcac tcacatatat tgatttatgt actgtttcca 4200 ttaaaaaagg taattcaaat gaaattgtta aatgtaatta attttgtttt cttgatgttt 4260 gtttcatcat cttcttttgc tcaggtaatt gaaatgaata attcgcctct gcgcgatttt 4320 gtaacttggt attcaaagca atcaggcgaa tccgttattg tttctcccga tgtaaaaggt 4380 actgttactg tatattcatc tgacgttaaa cctgaaaatc tacgcaattt ctttatttct 4440 gttttacgtg caaataattt tgatatggta ggttctaacc cttccattat tcagaagtat 4500 aatccaaaca atcaggatta tattgatgaa ttgccatcat ctgataatca ggaatatgat 4560 gataattccg ctccttctgg tggtttcttt gttccgcaaa atgataatgt tactcaaact 4620 tttaaaatta ataacgttcg ggcaaaggat ttaatacgag ttgtcgaatt gtttgtaaag 4680 tctaatactt ctaaatcctc aaatgtatta tctattgacg gctctaatct attagttgtt 4740 agtgctccta aagatatttt agataacctt cctcaattcc tttcaactgt tgatttgcca 4800 actgaccaga tattgattga gggtttgata tttgaggttc agcaaggtga tgctttagat 4860 ttttcatttg ctgctggctc tcagcgtggc actgttgcag gcggtgttaa tactgaccgc 4920 ctcacctctg ttttatcttc tgctggtggt tcgttcggta tttttaatgg cgatgtttta 4980 gggctatcag ttcgcgcatt aaagactaat agccattcaa aaatattgtc tgtgccacgt 5040 attcttacgc tttcaggtca gaagggttct atctctgttg gccagaatgt tccttttatt 5100 actggtcgtg tgactggtga atctgccaat gtaaataatc catttcagac gattgagcgt 5160 caaaatgtag gtatttccat gagcgttttt cctgttgcaa tggctggcgg taatattgtt 5220 ctggatatta ccagcaaggc cgatagtttg agttcttcta ctcaggcaag tgatgttatt 5280 actaatcaaa gaagtattgc tacaacggtt aatttgcgtg atggacagac tcttttactc 5340 ggtggcctca ctgattataa aaacacttct caggattctg gcgtaccgtt cctgtctaaa 5400 atccctttaa tcggcctcct gtttagctcc cgctctgatt ctaacgagga aagcacgtta 5460 tacgtgctcg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg 5520 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 5580 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 5640 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 5700 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 5760 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 5820 tatctcgggc tattcttttg atttataagg gattttgccg atttcggaac caccatcaaa 5880 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc 5940 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg 6000 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 6060 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct 6120 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat 6180 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc 6240 atgcctgcag gtcctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc 6300 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata 6360 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc 6420 gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc 6480 ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca 6540 tctacaccaa cgtgacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc 6600 cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga 6660 cgcgaattat ttttgatggc gttcctattg gttaaaaaat gagctgattt aacaaaaatt 6720 taatgcgaat tttaacaaaa tattaacgtt tacaatttaa atatttgctt atacaatctt 6780 cctgtttttg gggcttttct gattatcaac cggggtacat atgattgaca tgctagtttt 6840 acgattaccg ttcatcgatt ctcttgtttg ctccagactc tcaggcaatg acctgatagc 6900 ctttgtagat ctctcaaaaa tagctaccct ctccggcatt aatttatcag ctagaacggt 6960 tgaatatcat attgatggtg atttgactgt ctccggcctt tctcaccctt ttgaatcttt 7020 acctacacat tactcaggca ttgcatttaa aatatatgag ggttctaaaa atttttatcc 7080 ttgcgttgaa ataaaggctt ctcccgcaaa agtattacag ggtcataatg tttttggtac 7140 aaccgattta gctttatgct ctgaggcttt attgcttaat tttgctaatt ctttgccttg 7200 cctgtatgat ttattggatg tt 7222 <210> 14 <211> 41 <212> DNA <213> Artificial Sequence <220> <223> BamH I_SM_upper which is a primer used for site-directed mutation <400> 14 aaggccgctt ttgcgggatc ctcaccctca gcagcgaaag a 41 <210> 15 <211> 41 <212> DNA <213> Artificial Sequence <220> <223> BamH I_SM_lower which is a primer used for site-directed mutation <400> 15 tctttcgctg ctgagggtga ggatcccgca aaagcggcct t 41 <210> 16 <211> 90 <212> DNA <213> Artificial Sequence <220> <223> BamM13HK_P8_primer which is an extension primer used for          preparation <400> 16 ttaatggaaa cttcctcatg aaaaagtctt tagtcctcaa agcctctgta gccgttgcta 60 ccctcgttcc gatgctgtct ttcgctgctg 90 <210> 17 <211> 95 <212> DNA <213> Artificial Sequence <220> <223> M13HK_P8 which is a library oligonucleotide used for preparation <220> <221> misc_feature <222> (1) <223> n is a, g, c or t <220> <221> misc_feature <222> (1) <223> m is a or c <400> 17 aaggccgctt ttgcgggatc cnmnnmnnmnnmnnmn nmncagcagc gaaagacagc 60 atcggaacga gggtagcaac ggctacagag gcttt 95 <210> 18 <211> 50 <212> PRT <213> Artificial Sequence <220> <223> P8 protein of M13 phage <400> 18 Ala Glu Gly Asp Asp Pro Ala Lys Ala Ala Phe Asn Ser Leu Gln Ala   1 5 10 15 Ser Ala Thr Glu Tyr Ile Gly Tyr Ala Trp Ala Met Val Val Val Ile              20 25 30 Val Gly Ala Thr Ile Gly Ile Lys Leu Phe Lys Lys Phe Thr Ser Lys          35 40 45 Ala Ser      50

Claims (18)

A solution comprising a phage displaying a peptide or peptide capable of binding to a carbon material, and a colloidal solution comprising a carbon material, wherein said peptide or said phage is dispersed in said solution. delete delete The composition of claim 1, wherein the composition is printed on a substrate and then dried to form an electrode. 5. The composition of claim 4, wherein the bonding force between the carbon materials or the bonding force between the carbon material and the substrate is enhanced such that the carbon material exhibits increased stability in the aqueous solution. The composition according to claim 1, wherein the phage is a phage genetically engineered to have a binding capacity to a carbonaceous material. The method of claim 1, wherein the carbon material is selected from the group consisting of a graphene sheet, a highly oriented pyrolytic graphite (HOPG) sheet, a single-walled carbon nanotube, a double-walled carbon nanotube, A multi-walled carbon nanotube, and a fullerene. The composition of claim 1, wherein the phage displaying the peptide or peptide forms a network structure with the carbonaceous material. The composition according to claim 1, wherein the peptide comprises at least one selected from the group consisting of amino acid sequences of SEQ ID NOS: 1-12. The composition according to claim 1, wherein the phage is M13 phage, F1 phage, Fd phage, If1 phage, Ike phage, Zj / Z phage, Ff phage, Xf phage, Pf1 phage or Pf3 phage. The method of claim 1, wherein the phage is selected from the group consisting of the C-terminal of the peptide is linked to the N-terminus of the phage coat protein, or the peptide is inserted between consecutive amino acid sequences of the phage coat protein, Wherein the amino acid sequence is substituted. 12. The composition according to claim 11, wherein the envelope protein is at least one selected from the group consisting of p3, p6, p8 and p9 of M13 phage. A solution comprising a phage displaying a peptide or peptide having a binding ability to a carbon material, and a colloid solution containing a carbon material, wherein the peptide or the phage is dispersed in the solution. 14. The composition of claim 13, wherein the electrode is an electrochemical device. 14. The composition of claim 13, wherein the composition is an ink composition for inkjet printing. Ink-jet printing the composition for electrode printing of claim 13 on a substrate; And
And drying the printed composition to form an electrode. Ink-jet printing the composition for electrode printing of claim 13 on a substrate;
Drying the printed composition to form an electrode; And
And forming an enzyme on the formed electrode. 18. The method of claim 17, further comprising treating the polymeric material on the formed electrode prior to the step of forming the enzyme.

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