A biomimetic conductive polymer enzyme-based sensing material and a preparation method thereof

Abstract

This invention relates to a biomimetic conductive polymeric enzyme-based sensing material and its preparation method. The biomimetic conductive polymer contains zwitterionic groups and redox groups, and when coated with a biological enzyme, it can form an enzyme-based sensing material that resists non-specific adhesion, maintains enzyme activity for a long time, and is oxygen-independent. The method involves first dissolving the comonomer and dispersing the biological enzyme using an ionic liquid, followed by electrochemical polymerization deposition to obtain the biomimetic conductive polymeric enzyme-based sensing material. This preparation method can electrochemically copolymerize conductive polymeric monomers of different polarities or solubilities, and is suitable for preparing multi-component copolymerized conductive polymeric sensing materials. The impedance of this biomimetic conductive polymeric enzyme-based sensing material is 1 Ω·cm. ‑2 ~10Ω·cm ‑2 It has a detection limit greater than 2 μM, and the relative enzyme activity is 88%–100%, which is 80%–90% after 4 weeks. It is suitable for various biosensors, including enzyme-based sensors, and can achieve highly sensitive detection in complex biological environments while maintaining the long-term stability of bioactive substances. It has broad application value.

Description

Technical Field

[0001] This invention relates to a biomimetic conductive polymer enzyme-based sensing material with both anti-nonspecific adhesion and high charge transport efficiency, and its preparation method, belonging to the field of conductive polymer enzyme-based sensing materials. Background Technology

[0002] Enzyme biosensors are devices that immobilize biological enzymes onto electrode surfaces as molecular recognition elements. These enzymes react with analytes via enzymatic reactions, and the detection signal is characterized by current or potential. Enzyme biosensors offer high selectivity, high sensitivity, rapid response, and small size, enabling real-time online monitoring. Therefore, enzyme sensors occupy an important position in wearable devices, environmental monitoring, food analysis, and biomedicine.

[0003] Compared with other materials, conductive polymers such as polypyrrole (PPy), polythiophene (PTh), and poly(3,4-ethylenedioxythiophene) (PEDOT) exhibit better flexibility and biocompatibility, excellent ionic / electronic conductivity, and low interfacial impedance, making them ideal biosensor materials. Furthermore, conductive polymers possess abundant porous structures, providing as many interfaces as possible to facilitate charge transfer between enzymes and materials. CN114563453A describes the fabrication of an enzyme biosensor by immobilizing horseradish catalase within a PEDOT carrier via electrochemical polymerization; the introduction of PEDOT effectively improves the conductivity of the enzyme biosensor. CN104280441A provides a method for immobilizing glucose oxidase-modified electrodes based on Au@Ag-PPy composite nanomaterials and L-cysteine ​​bilayer membranes; this electrode has a glucose detection range of 2.5 × 10⁻⁶. -7 ~1.2*10 -4 The addition of mol / L PPy improved the sensing sensitivity and broadened the linear range; KR2020084678A prepared a glucose sensor by combining PTh with chitosan-immobilized glucose oxidase using a layer-by-layer assembly method. The introduction of PTh improved the ionic / electronic conductivity and reduced the detection limit of the sensor.

[0004] While the aforementioned technologies partially address the low sensitivity issue of enzyme-based sensors by encapsulating enzymes in conductive polymer materials, thus meeting the detection requirements for some biomarkers, they still struggle to meet detection standards for low-abundance biomarkers such as adrenaline. Furthermore, electrolyte solutions and electrochemical conditions are not conducive to maintaining the enzyme's three-dimensional structure and activity. Additionally, enzyme-based sensors operate in complex biological environments such as sweat, tissue fluid, and blood. Biomolecules (proteins, cells, and microorganisms, etc.) can form a protein adhesion layer on the conductive polymer electrodes through non-specific adhesion, weakening the biosensor's performance, resulting in poor signal-to-noise ratio and high background signal, severely impacting the sensor's sensitivity, repeatability, and reliability. Simultaneously, the strong hydrophobic-hydrophobic interactions between conductive polymer materials and enzymes easily lead to enzyme inactivation, significantly affecting the long-term storage and performance of enzyme-based sensors. Summary of the Invention

[0005] To address the issues of non-specific adsorption weakening sensing performance and enzyme inactivation in conductive polymeric enzyme-based sensing materials, the inventors, through extensive and systematic research, discovered that zwitterions, due to their superhydrophilic and electroneutrally neutral properties, can effectively resist non-specific adsorption by proteins and cells in complex biological environments, ensuring the stability of enzyme sensors. Simultaneously, the encapsulation of enzymes in zwitterionic conductive polymers provides a hydrophilic environment, effectively inhibiting hydrophobic-hydrophobic interactions between the original conductive polymer and the enzyme, significantly improving the long-term storage and operational stability of enzyme-based sensors. Furthermore, considering the insufficient sensitivity of conductive polymeric enzyme-based sensing materials, the inventors, through extensive innovative work, synthesized and developed a series of copolymerized conductive polymers combining redox functionalized monomers and zwitterionic functionalized monomers. Extensive experimental verification has shown that these copolymerized conductive polymers not only maintain the resistance of the original zwitterionic conductive polymers to non-specific protein adsorption and enzyme activity, but also significantly improve sensing sensitivity by utilizing the volume effect of the electron mediating effect of the redox functionalized conductive polymers, thus comprehensively solving multiple problems of existing conductive polymeric enzyme-based sensing materials. The main problems with modifying conductive polymers with redox groups are: they significantly distort the polymer chains, reducing the overlap of conjugated orbitals and thus degrading the conductivity of the molecular chains; simultaneously, side chain modification increases the distance between molecular chains, making electronic transitions between chains difficult; although redox groups can increase the conductivity of materials at oxidation potential, they also hinder inter-chain conductivity without an applied charge. Therefore, those skilled in the art of conductive polymers generally believe that redox group modification of conductive polymers will weaken conductivity. However, the invention unexpectedly discovered that by introducing redox groups with suitable structures and using optimized linking chemical structures, redox group-functionalized conductive polymers can be endowed with extremely low interfacial impedance, further significantly improving the sensitivity of enzyme sensing.

[0006] The inventors prepared conductive polymeric enzyme-based sensing materials using electrochemical in-situ polymerization encapsulation. This method involves dissolving the enzyme and polymer monomers in the same solvent, and then encapsulating the enzyme within a continuously growing polymer network using electrochemical methods such as galvanostatics, galvanostatics, or cyclic voltammetry. The advantages of this method are its simplicity, the one-step polymerization and encapsulation process, and the controllable thickness of the sensing material. However, the inventors discovered that zwitterionic functionalized monomers are extremely hydrophilic, making it difficult to electrochemically copolymerize with redox functionalized monomers in the same solvent, thus significantly limiting the application of this copolymer in enzyme-based sensing materials. Through extensive creative work, the inventors also developed a surfactant-assisted electrochemical polymerization method to prepare zwitterionic functionalized conductive polymer materials. However, experimental results showed that surfactants can damage enzymes, proteins, and some highly chemically active chemical groups. While utilizing the strong solubility of ionic liquids to prepare high-concentration electrolyte solutions has the potential to solve the problems of dissolution and electrochemical copolymerization of monomers with similar polarity involved in this invention, both ionic liquids and electrolytes are composed of anions and cations, which may strongly interact with the amino and carboxyl groups expressed in large quantities on the enzyme surface, thereby causing enzyme denaturation and loss of activity. For the above reasons, those skilled in the art of electrochemistry generally believe that ionic liquids containing high-concentration electrolytes are unlikely to maintain the three-dimensional structure and activity of enzymes. However, unexpectedly, the inventors discovered that by using suitable ionic liquids, suitable electrolytes, and introducing trace amounts of water, under optimized electrochemical conditions, conductive polymeric enzyme-based sensing materials with high enzyme activity can be prepared through electrochemical polymerization. Furthermore, given that zwitterionic monomers and redox functionalized monomers have very different oxidation potentials in ionic liquid electrolyte solutions, those skilled in the art also believe that it is difficult to effectively control the composition of the deposited film by changing the composition of the monomers in the electrolyte solution. However, this invention further regulates the structure of the ionic liquid and electrolyte, changes their interaction with the monomers, and further optimizes the electrochemical conditions based on the difference in oxidation barriers of the two monomers, thus successfully developing an effective method for the controllable preparation of the above-mentioned conductive polymeric enzyme-based sensing materials based on ionic liquid electrolytes.

[0007] The present invention has three objectives. First, it provides a copolymer conductive polymer that simultaneously possesses resistance to nonspecific adhesion, volumetric redox capabilities, and ultra-low impedance, consisting of both zwitterionic functionalized monomers and redox functionalized monomers. Second, it utilizes the aforementioned conductive copolymer to encapsulate enzymes to prepare conductive polymeric enzyme-based sensing materials. By leveraging its excellent resistance to nonspecific protein adhesion, volumetric redox properties, and low impedance, the conductive polymeric enzyme-based sensing materials achieve long-term stability of enzyme activity, operational stability in complex biological environments, and high-sensitivity detection. Third, it provides a method for preparing this conductive copolymer through electrochemical copolymerization in ionic liquids and then encapsulating biological enzymes to prepare high-enzyme-activity enzyme biosensing materials. This effectively solves the problems of co-dissolution of multiple monomers with large polarity differences and the damage to the activity of enzymes and functionalized groups caused by the original electrochemical method.

[0008] To achieve the objectives of this invention, the following solution is adopted:

[0009] A biomimetic conductive polymer enzyme-based sensing material possessing resistance to nonspecific adhesion, volume-effect redox capability, and ultra-low impedance, characterized by comprising a monomer unit of formula (I):

[0010]

[0011] In the general structural formula (Ⅰ):

[0012] These are groups that form the backbone of the conductive polymer.

[0013] -L1- is -(CH2) x -、-(CH2) x -O-(CH2) y -、-(CH2) x -O-CO-(CH2) y -、-(CH2) x -(CH2-CH2-O) y -、-(CH2) x -O-(CH2-CH2-O) y -、-O-(CH2) x -(CH2-CH2-O) y -or -O-(CH2) x -O-(CH2-CH2-O) y - any one of the following, where x and y are integers, and 0 ≤ x ≤ 20, 0 ≤ y ≤ 20.

[0014] -L2- is -(CH2) x -、-O-(CH2) x -、-C(O)-O-(CH2) x-、-C(O)-NH-(CH2) x -、-C(CH3)2-C(O)-O-(CH2) x -、-CH(CH3)-O-(CH2) x -、-S-CH2-CH2-O-(CH2) x -, -S-CH2-CH(CH3)-C(O)-O-(CH2) x - any one or more of the following, where x is an integer and 0 ≤ x ≤ 20.

[0015] A biomimetic conductive polymer enzyme-based sensing material possessing resistance to nonspecific adhesion, volume-effect redox capability, and ultra-low impedance is characterized by comprising, in addition to the monomer unit of formula (I), a monomer unit as shown in formula (II):

[0016]

[0017] In the general structural formula (II):

[0018] These are groups that form the backbone of the conductive polymer.

[0019] -L3- can be any one or more of -CH2-, -O-, -S-, -NH-, -C(O)-, -C(O)NH-, -NH-C(O)-, -OC(O)-, or -C(O)O-;

[0020] -L4- is -(CH2) x -、-(CH2) x -O-(CH2) y -、-(CH2) x -O-CO-(CH2) y -、-(CH2) x -(CH2-CH2-O) y -、-(CH2) x -O-(CH2-CH2-O) y -、-O-(CH2) x -(CH2-CH2-O) y -or -O-(CH2) x -O-(CH2-CH2-O) y - any one of the following, where x and y are integers, and 0 ≤ x ≤ 20, 0 ≤ y ≤ 20;

[0021] -R2 is a redox functionalized group.

[0022] As a preferred technical solution:

[0023] The aforementioned single unit Preferred Any one of its derivatives.

[0024] In the aforementioned monomer unit structure formula (I), -L1- is preferably -(CH2). x -、-(CH2) x -O-(CH2) y -、-(CH2) x -O-CO-(CH2) y -、-(CH2) x -(CH2-CH2-O) y - any one of the following, where x and y are integers, and 0 ≤ x ≤ 5, 0 ≤ y ≤ 5.

[0025] In the aforementioned monomer unit structure formula (I), -L2- is preferably -(CH2). x -、-O-(CH2) x -、-C(O)-O-(CH2) x -、-C(O)-NH-(CH2) x -、-C(CH3)2-C(O)-O-(CH2) x -、-CH(CH3)-O-(CH2) x - any one or more of the following; where x and y are integers, and 0 ≤ x ≤ 5, 0 ≤ y ≤ 5.

[0026] In the general formula (Ⅱ) of the monomer unit structure, -L3- is preferably any one of -CH2-, -O-, -S-, -NH-, -C(O)-, -NH-C(O)-, and -OC(O)-.

[0027] In the aforementioned monomer unit structure formula (II), -L4- is preferably -(CH2). x -、-(CH2) x -O-(CH2) y -、-(CH2) x -O-CO-(CH2) y -、-(CH2) x -(CH2-CH2-O) y -、-O-(CH2) x -(CH2-CH2-O) y - any one of the following, where x and y are integers, and 0 ≤ x ≤ 20, 0 ≤ y ≤ 20.

[0028] In the aforementioned single-unit, -R1 is Any one of them.

[0029] In the aforementioned single-unit, -R2 is preferably... Any one of them.

[0030] The prepared copolymer conductive polymer is characterized by electrochemical polymerization of one or more formulas (I) and one or more formulas (II) in a certain proportion. The polymer has the following general structural formula (III):

[0031]

[0032] Where {a1,a2,a3,……,ap} are integers, and 0≤{a1,a2,a3,……,ap}≤100; {b1,b2,b3,……,bq} are integers, and 0≤{b1,b2,b3,……,bq}≤1000.

[0033] The method for preparing a polymer-based enzyme sensing material as described above involves electrochemical copolymerization of the aforementioned conductive polymer to encapsulate a biological enzyme in an ionic liquid, and includes the following steps:

[0034] (1) The 0mM~50mM conductive polymer monomer M 11 M 12 M 13 ...M 1p One or more of the following, 0mM to 50mM conductive polymer monomers M 21 M 22 M 23 ...M 2q One or more of the following, biological enzymes, trace amounts of water, and electrolytes are added to the ionic liquid and stirred until dissolved.

[0035] (2) Add the solution to the electrolytic cell and use a three-electrode system. Set up the working electrode, Ag / AgCl reference electrode and platinum mesh counter electrode. The enzyme concentration range is 0.5 mg / mL to 15 mg / mL, the water content is 0.01% to 15%, the electrolyte concentration is 0 mM to 500 mM, and the polymerization potential range is -1 V to 3 V. The electrochemical enzyme sensing electrode is obtained.

[0036] The polymer-based enzyme sensing material described above may contain a bioenzyme selected from at least one of glucose oxidase, glucose dehydrogenase, galactose oxidase, cholesterol oxidase, amino acid oxidase, alcohol oxidase, lactate oxidase, laccase, catalase, and uricase.

[0037] The electrolyte is at least one of lithium chloride, lithium perchlorate, tetrabutylammonium perchlorate, tetrabutylammonium hexafluorophosphate, tetrabutylammonium tetrafluoroborate, lithium dioxaborate, and lithium bis(trifluoromethanesulfonyl)imide.

[0038] The ionic liquid is characterized in that it is composed of cations and anions, and has A + B - The general structural formula, where A + Represents a cation, for One type; where -R1, -R2, -R3, and -R4 are -H and -(CH2). x -、-(CH3) x -(CH2) x CH3, -C x H 2x+1 N≡C(CH2) x -, -CH=CH-(CH3) x -CH2-CH2-(O-CH2-CH2) x -OH or -CH2-CH2-(O-CH2-CH-(CH3)x) y One or more of the -OH groups. Where x and y are integers, and 0≤x≤30, 0≤y≤30.

[0039] The B mentioned above - It is an anion, and is Br. - Cl - I - F - BF4 - PF4 - PF6 - oTf - ,HAc - HSO4 - EtSO4 - SCN - CN - OCN - CNO - CF3SO3 - CF3COO - (CF3SO2)2N - (CF3SO2)2Cl - (HO)2PO2 - AcO - One of them.

[0040] As a preferred technical solution:

[0041] The bioenzyme is preferably at least one of glucose oxidase, lactate oxidase, and laccase.

[0042] The electrolyte is selected based on its solubility and doping effect on electrochemical polymerization, and is preferably at least one of lithium perchlorate, tetrabutylammonium perchlorate, and tetrabutylammonium hexafluorophosphate.

[0043] The ionic liquid is selected based on its solubility in various monomers, its ability to maintain enzyme activity, and its controllability in electrochemical polymerization. The preferred ionic liquids are the following six types:

[0044] The method for preparing an enzyme sensor by electrochemical copolymerization of conductive polymer-embedded biological enzymes in ionic liquids preferably includes an enzyme concentration range of 1 mg / mL to 10 mg / mL; a polymerization potential range of -0.8 V to 1.4 V; and an electrolyte concentration of 10 mM to 100 mM. Since increasing the water content is detrimental to the electrochemical polymerization effect but beneficial to the maintenance of enzyme activity, the water content is preferably 0.1% to 5%.

[0045] Principle of this invention:

[0046] In the sensing material designed and developed in this invention, structure (I) contains zwitterionic groups, which can effectively resist the non-specific adsorption of proteins and cells in complex biological environments by utilizing their superhydrophilic and electroneutrally neutral properties, thus ensuring the stability of enzyme sensors. At the same time, the zwitterionic conductive polymer material encapsulating enzymes can provide a hydrophilic environment, effectively inhibiting the hydrophobic-hydrophobic interaction between the original conductive polymer material and the enzyme, and significantly improving the long-term storage and stability of enzyme-based sensors. Structure (II) contains redox groups, which can utilize the volume effect of their electron mediating effect to endow the sensing material with very low interfacial impedance, thus significantly improving sensing sensitivity. This invention utilizes the superior solubility of ionic liquids to solve the problem of co-dissolution of monomers with dissimilar polarities. By employing suitable zwitterionic and electrolyte structures and trace water content, it overcomes the damage to enzyme activity caused by anions and cations, as well as the differences in oxidation barriers of comonomers. An effective method for controllably preparing the aforementioned conductive polymeric enzyme-based sensing materials has been developed, resulting in a copolymeric conductive polymer and its enzyme-based sensing material that simultaneously possesses resistance to nonspecific adhesion, volume-effect redox capabilities, and ultra-low impedance. This achieves long-term stability of enzyme activity, operational stability in complex biological environments, and high-sensitivity detection of the conductive polymeric enzyme-based sensing material.

[0047] Beneficial effects

[0048] 1. The present invention provides a copolymer conductive polymer sensing material that simultaneously possesses resistance to nonspecific adhesion, volume effect redox capability, and ultra-low impedance. It can be widely used in various biosensors, including enzyme-based sensors, to maintain the long-term stability of bioactive substances while achieving highly sensitive detection in complex biological environments.

[0049] 2. The present invention provides a method for preparing copolymer conductive polymer sensing materials with anti-nonspecific adhesion, volume effect redox capability, and ultra-low impedance based on ionic liquid electrochemistry. This method can be applied to electrochemical copolymerization of conductive polymer monomers with different polarities and solubilities. It is not only applicable to binary copolymerization, but also realizes the idea of ​​multi-component copolymerization to endow materials with more properties. Attached Figure Description

[0050] Figure 1 The electrochemical copolymerization curve is shown for the copolymerase electrode prepared in Example 1 of this invention.

[0051] Figure 2 The homogenization response of the copolymerase electrode prepared in Example 1 of this invention to glucose after different storage times is shown. Detailed Implementation

[0052] To make the present invention more apparent and understandable, preferred embodiments are described in detail below:

[0053] Example 1

[0054] In this embodiment, electrochemical copolymerization was employed. 31.92 mg of lithium perchlorate (LiClO4), 2.01 mg of monomer I-1, and 10.2 mg of monomer II-1 were added to 3 mL of 1-ethyl-3-methylimidazolium tetrafluoroborate ([EMIM][BF4]) and stirred until dissolved. 50 μL of an 8 mg / mL glucose oxidase (GOx) aqueous solution was drawn and mixed thoroughly to obtain a mixed solution containing 1.6% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.4 V to +1.3 V, thus obtaining the copolymer enzyme electrode. Figure 1 The electrochemical copolymerization curves of the copolymer enzyme electrode prepared in this embodiment are shown. With the increase in the number of polymerization cycles, the oxidation peak and peak current of the PEDOT-PC polymer show a significant upward trend, proving that the thin film material is continuously deposited on the gold substrate surface; after GOx composite, the enzyme activity remains at 100%; after electrochemical polymerization, the relative enzyme activity of GOx remains at 100% as shown by UV absorption spectroscopy; with the increase of the EDOT-PC feed ratio, the anti-protein adhesion performance is better; the QCM chip frequency decreases by 3 Hz; with the increase of the EDOT-Fc feed ratio, the charge transfer impedance decreases accordingly, from 102.36 Ω·cm. -2 Reduced to 5.91 Ω·cm -2 The glucose sensing limit is 10 μM. Figure 2 The homogenization response of the copolymerase electrode prepared in this embodiment to glucose after different storage times shows that it can maintain 90% of the response after 4 weeks.

[0055] Example 2

[0056] In this embodiment, electrochemical copolymerization was employed. 69.15 mg of tetrabutylammonium perchlorate (TBAP), 2.01 mg of monomer I-2, and 10.2 mg of monomer II-2 were added to 3 mL of 1-ethyl-3-methylimidazolium tetrafluoroborate ([EMIM][BF4]) and stirred until dissolved. 50 μL of an 8 mg / mL glucose oxidase (GOx) aqueous solution was drawn and mixed thoroughly to obtain a mixed solution containing 1.6% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.4 to +1.3 V, thus obtaining the copolymer enzyme electrode. After GOx was incorporated, the enzyme activity remained at 100%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of GOx remained at 99.4%. At this copolymerization ratio, the QCM chip frequency decreased by 5 Hz, and the charge transfer impedance was 5.35 Ω·cm. -2 The sensing limit for glucose is 13 μM, and it retains 89% of its response after 4 weeks.

[0057] Example 3

[0058] In this embodiment, electrochemical copolymerization was employed. 73.28 mg of tetrabutylammonium hexafluorophosphate (TBHAP), 2.01 mg of I-2 monomer, and 10.2 mg of II-2 monomer were added to 3 mL of 1-ethyl-3-methylimidazolium tetrafluoroborate ([EMIM][BF4]) and stirred until dissolved. 50 μL of an 8 mg / mL glucose oxidase (GOx) aqueous solution was drawn and mixed thoroughly to obtain a mixed solution containing 1.6% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.4 to +1.3 V, thus obtaining the copolymer enzyme electrode. After GOx was incorporated, the enzyme activity remained at 100%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of GOx remained at 99.5%. At this copolymerization ratio, the QCM chip frequency decreased by 6 Hz, and the charge transfer impedance was 5.06 Ω·cm. -2 The sensing limit for glucose is 15 μM, and it retains 88.6% of its response after 4 weeks.

[0059] Example 4

[0060] In this embodiment, electrochemical copolymerization was employed. 29.16 mg of lithium perchlorate (LiClO4), 2.01 mg of I-2 monomer, and 10.2 mg of II-2 monomer were added to 3 mL of 1-ethyl-3-methylimidazolium tetrafluoroborate ([EMIM][BF4]) and stirred until dissolved. 50 μL of an 8 mg / mL glucose oxidase (GOx) aqueous solution was drawn and mixed thoroughly to obtain a mixed solution containing 1.6% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.4 to +1.2 V, thus obtaining the copolymer enzyme electrode. After GOx was incorporated, the enzyme activity remained at 100%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of GOx remained at 99.6%. At this copolymerization ratio, the QCM chip frequency decreased by 3.5 Hz, and the charge transfer impedance was 5.77 Ω·cm. -2 The sensing limit for glucose is 10 μM, and it retains 89.9% of its response after 4 weeks.

[0061] Example 5

[0062] In this embodiment, electrochemical copolymerization was employed. 29.15 mg of tetrabutylammonium perchlorate (TBAP), 4.15 mg of monomer I-2, and 5.32 mg of monomer II-2 were added to 3 mL of 1-ethyl-3-methylimidazolium trifluoromethanesulfonate ([EMIM][oTf]) and stirred until dissolved. 60 μL of a 9 mg / mL aqueous solution of lactate oxidase (LOx) was drawn and mixed thoroughly to obtain a mixed solution containing 2.0% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.5 V to +1.2 V, thus obtaining the copolymer enzyme electrode. After LOx was incorporated, the enzyme activity remained above 95%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of LOx remained at 94.2%. At this copolymerization ratio, the QCM chip frequency decreased by 10 Hz, and the charge transfer impedance was 3.38 Ω·cm. -2 The sensing limit for lactic acid is 10 μM, and it retains 87% of its response after 4 weeks.

[0063] Example 6

[0064] In this embodiment, electrochemical copolymerization was employed. 15.87 mg of tetrabutylammonium hexafluorophosphate (TBHAP), 1.52 mg of I-3 monomer, and 13.6 mg of II-3 monomer were added to 3 mL of 1-butyl-1-methylpyrrolidine trifluoromethanesulfonate ([C4Clpy][oTf]) and stirred until dissolved. 63 μL of an 8 mg / mL laccase aqueous solution was then drawn and mixed thoroughly to obtain a mixed solution containing 2.1% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.6 V to +1.1 V, thus obtaining the copolymer enzyme electrode. After lacase conjugation, the enzyme activity remained above 95%; UV absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of lacase remained at 94.4%. At this copolymerization ratio, the QCM chip frequency drops by 15 Hz, and the charge transfer impedance is 2.29 Ω·cm. -2 The sensing limit for adrenaline is 2 μM, and it retains 85% of its response after 4 weeks.

[0065] Example 7

[0066] In this embodiment, electrochemical copolymerization was employed. 106.2 mg of lithium perchlorate (LiClO4), 2.01 mg of I-4 monomer, and 10.2 mg of II-4 monomer were added to 3 mL of 1-ethyl-3-methylimidazolium hydrochloride ([EMIM][AC]) and stirred until dissolved. 66 μL of a 7 mg / mL glucose oxidase (GOx) aqueous solution was drawn and mixed thoroughly to obtain a mixed solution containing 2.2% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.7 V to +1.0 V, thus obtaining the copolymer enzyme electrode. After GOx was incorporated, the enzyme activity remained above 95%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of GOx remained at 94.1%. At this copolymerization ratio, the QCM chip frequency decreased by 28 Hz, and the charge transfer impedance was 1.05 Ω·cm. -2 It has a glucose sensing limit of 30 μM and can maintain 80% of its response after 4 weeks.

[0067] Example 8

[0068] In this embodiment, electrochemical copolymerization was employed. 53.15 mg of tetrabutylammonium perchlorate (TBAP), 4.61 mg of monomer I-5, and 7.89 mg of monomer II-5 were added to 3 mL of 1-ethyl-3-methylimidazolium sulfate ([EMIM][EtSO4]) and stirred until dissolved. 69 μL of a 6 mg / mL aqueous solution of lactate oxidase (LOx) was drawn and mixed thoroughly to obtain a mixed solution containing 2.3% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.8 V to +1.4 V, thus obtaining the copolymer enzyme electrode. After LOx was incorporated, the enzyme activity remained above 92%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of LOx remained at 91.3%. At this copolymerization ratio, the QCM chip frequency decreased by 1 Hz, and the charge transfer impedance was 6.35 Ω·cm. -2 The sensing limit for lactic acid is 15 μM, and it retains 82% of its response after 4 weeks.

[0069] Example 9

[0070] In this embodiment, electrochemical copolymerization was employed. 28.73 mg of tetrabutylammonium hexafluorophosphate (TBHAP), 5.8 mg of I-6 monomer, and 6.79 mg of II-6 monomer were added to 3 mL of 1-ethyl-3-methylimidazolium hydrogen sulfate ([EMIM][HSO4]) and stirred until dissolved. 72 μL of a 5 mg / mL laccase aqueous solution was drawn and mixed thoroughly to obtain a mixed solution containing 2.4% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.3 V to +1.5 V, thus obtaining the copolymer enzyme electrode. After lacase conjugation, the enzyme activity remained above 95%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of lacase remained at 94.6%. At this copolymerization ratio, the QCM chip frequency decreased by 2 Hz, and the charge transfer impedance was 7.36 Ω·cm. -2 The sensing limit for adrenaline is 5 μM, and it retains 80% of its response after 4 weeks.

[0071] Example 10

[0072] In this embodiment, electrochemical copolymerization was employed. 112.6 mg of lithium perchlorate (LiClO4), 6.71 mg of I-7 monomer, and 13.2 mg of II-7 monomer were added to 3 mL of 1-ethyl-3-methylimidazolium tetrafluoroborate ([EMIM][BF4]) and stirred until dissolved. 78 μL of a 4 mg / mL glucose oxidase (GOx) aqueous solution was drawn and mixed thoroughly to obtain a mixed solution containing 2.6% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.4 V to +1.15 V, thus obtaining the copolymer enzyme electrode. After GOx was incorporated, the enzyme activity remained above 90%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of GOx remained at 89.7%. At this copolymerization ratio, the QCM chip frequency decreased by 8 Hz, and the charge transfer impedance was 6.37 Ω·cm. -2 The sensing limit for glucose is 25 μM, and it retains 82% of its response after 4 weeks.

[0073] Example 11

[0074] In this embodiment, electrochemical copolymerization was employed. 56.75 mg of tetrabutylperchloramine (TBAP), 3.89 mg of I-8 monomer, and 10.6 mg of II-8 monomer were added to 3 mL of 1-ethyl-3-methylimidazolium trifluoromethanesulfonate ([EMIM][oTf]) and stirred until dissolved. 84 μL of a 2 mg / mL aqueous solution of lactate oxidase (LOx) was drawn and mixed thoroughly to obtain a mixed solution containing 2.8% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.5 V to +1.25 V, thus obtaining the copolymer enzyme electrode. After LOx was incorporated, the enzyme activity remained above 90%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of LOx remained at 89.2%. At this copolymerization ratio, the QCM chip frequency decreased by 17 Hz, and the charge transfer impedance was 2.38 Ω·cm. -2 The sensing limit for lactic acid is 20 μM, and it retains 83% of its response after 4 weeks.

[0075] Example 12

[0076] In this embodiment, electrochemical copolymerization was employed. 31.78 mg of tetrabutylammonium hexafluorophosphate (TBHAP), 8.12 mg of I-9 monomer, and 6.3 mg of II-9 monomer were added to 3 mL of 1-butyl-1-methylpyrrolidine trifluoromethanesulfonate ([C4Clpy][oTf]) and stirred until dissolved. 93 μL of a 2 mg / mL laccase aqueous solution was then drawn and mixed thoroughly to obtain a mixed solution containing 3.1% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.6 V to +1.35 V, thus obtaining the copolymer enzyme electrode. After lacase conjugation, the enzyme activity remained above 90%; UV absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of lacase remained at 89.6%. At this copolymerization ratio, the QCM chip frequency drops by 25 Hz, and the charge transfer impedance is 1.09 Ω·cm. -2 The sensing limit for adrenaline is 10 μM, and it retains 81% of its response after 4 weeks.

[0077] Example 13

[0078] In this embodiment, electrochemical copolymerization was employed. 31.92 mg of lithium perchlorate (LiClO4), 3.41 mg of I-10 monomer, and 15.22 mg of II-10 monomer were added to 3 mL of 1-ethyl-3-methylimidazolium hydrochloride ([EMIM][AC]) and stirred until dissolved. 102 μL of a 1 mg / mL glucose oxidase (GOx) aqueous solution was drawn and mixed thoroughly to obtain a mixed solution containing 3.4% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.8 V to +1.45 V, thus obtaining the copolymer enzyme electrode. After GOx was incorporated, the enzyme activity remained above 90%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of GOx remained at 88.9%. At this copolymerization ratio, the QCM chip frequency decreased by 1 Hz, and the charge transfer impedance was 8.40 Ω·cm. -2 The sensing limit for glucose is 30 μM, and it retains 82% of its response after 4 weeks.

[0079] Example 14

[0080] In this embodiment, electrochemical copolymerization was employed. 50.98 mg of tetrabutylammonium perchlorate (TBAP), 4.72 mg of I-11 monomer, and 11.7 mg of II-11 monomer were added to 3 mL of 1-ethyl-3-methylimidazolium sulfate ethyl ester ([EMIM][EtSO4]) and stirred until dissolved. 105 μL of a 2 mg / mL lactate oxidase (LOx) aqueous solution was extracted and mixed thoroughly to obtain a mixed solution containing 3.5% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.8 V to +1.15 V, thus obtaining the copolymer enzyme electrode. After LOx was incorporated, the enzyme activity remained above 90%; UV absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of LOx remained at 89.8%. At this copolymerization ratio, the QCM chip frequency decreased by 3 Hz, and the charge transfer impedance was 6.41 Ω·cm. -2 The sensing limit for lactic acid is 30 μM, and it retains 82% of its response after 4 weeks.

[0081] Example 15

[0082] In this embodiment, electrochemical copolymerization was employed. 72.8 mg of tetrabutylammonium hexafluorophosphate (TBHAP), 7.31 mg of I-12 monomer, and 9.89 mg of II-12 monomer were added to 3 mL of 1-ethyl-3-methylimidazolium hydrogen sulfate ([EMIM][HSO4]) and stirred until dissolved. 12 μL of a 3 mg / mL laccase aqueous solution was then drawn and mixed thoroughly to obtain a mixed solution containing 0.4% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.1 V to +1.2 V, thus obtaining the copolymer enzyme electrode. After lacase conjugation, the enzyme activity remained above 90%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of lacase remained at 88.7%. At this copolymerization ratio, the QCM chip frequency decreased by 11 Hz, and the charge transfer impedance was 4.42 Ω·cm. -2 The sensing limit for adrenaline is 15 μM, and it retains 82% of its response after 4 weeks.

[0083] Example 16

[0084] In this embodiment, electrochemical copolymerization was employed. 56.2 mg of lithium perchlorate (LiClO4), 8.8 mg of I-13 monomer, and 7.34 mg of II-13 monomer were added to 3 mL of 1-ethyl-3-methylimidazolium tetrafluoroborate ([EMIM][BF4]) and stirred until dissolved. 24 μL of a 5 mg / mL glucose oxidase (GOx) aqueous solution was extracted and mixed thoroughly to obtain a mixed solution containing 0.8% water. 4 mL of this mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.2 V to +1.25 V, thus obtaining the copolymer enzyme electrode. After GOx was incorporated, the enzyme activity remained above 90%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of GOx remained at 89.1%. At this copolymerization ratio, the QCM chip frequency decreased by 14 Hz, and the charge transfer impedance was 2.43 Ω·cm. -2 The sensing limit for glucose is 25 μM, and it retains 83% of its response after 4 weeks.

[0085] Example 17

[0086] In this embodiment, electrochemical copolymerization was employed. 32.7 mg of tetrabutylammonium perchlorate (TBAP), 10.77 mg of I-14 monomer, and 5.63 mg of II-14 monomer were added to 3 mL of 1-ethyl-3-methylimidazolium trifluoromethanesulfonate ([EMIM][oTf]) and stirred until dissolved. 132 μL of a 5 mg / mL aqueous solution of lactate oxidase (LOx) was drawn off and mixed thoroughly to obtain a mixed solution containing 4.4% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.3 V to +1.3 V, thus obtaining the copolymer enzyme electrode. After LOx was incorporated, the enzyme activity remained above 90%; UV absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of LOx remained at 88.9%. At this copolymerization ratio, the QCM chip frequency decreased by 26 Hz, and the charge transfer impedance was 1.34 Ω·cm. -2 The sensing limit for lactic acid is 30 μM, and it retains 83% of its response after 4 weeks.

[0087] Example 18

[0088] In this embodiment, electrochemical copolymerization was employed. 21.5 mg of tetrabutylammonium hexafluorophosphate (TBHAP), 5.18 mg of I-15 monomer, and 9.3 mg of II-15 monomer were added to 3 mL of 1-butyl-1-methylpyrrolidine trifluoromethanesulfonate ([C4Clpy][oTf]) and stirred until dissolved. 135 μL of a 6 mg / mL laccase aqueous solution was then drawn and mixed thoroughly to obtain a mixed solution containing 4.5% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.4 V to +1.35 V, thus obtaining the copolymer enzyme electrode. After lacase conjugation, the enzyme activity remained above 90%; UV absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of lacase remained at 89.3%. At this copolymerization ratio, the QCM chip frequency decreased by 1.5 Hz, and the charge transfer impedance was 8.15 Ω·cm. -2 The sensing limit for adrenaline is 13 μM, and it retains 81% of its response after 4 weeks.

[0089] Example 19

[0090] In this embodiment, electrochemical copolymerization was employed. 76.1 mg of lithium perchlorate (LiClO4), 8.09 mg of I-16 monomer, and 7.94 mg of II-16 monomer were added to 3 mL of 1-ethyl-3-methylimidazolium hydrochloride ([EMIM][AC]) and stirred until dissolved. 144 μL of a 7 mg / mL glucose oxidase (GOx) aqueous solution was extracted and mixed thoroughly to obtain a mixed solution containing 4.8% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.5 V to +1.4 V, thus obtaining the copolymer enzyme electrode. After GOx was incorporated, the enzyme activity remained above 90%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of GOx remained at 88.6%. At this copolymerization ratio, the QCM chip frequency decreased by 1.8 Hz, and the charge transfer impedance was 7.36 Ω·cm. -2 The sensing limit for glucose is 25 μM, and it retains 83% of its response after 4 weeks.

[0091] Example 20

[0092] In this embodiment, electrochemical copolymerization was employed. 32.05 mg tetrabutylammonium perchlorate (TBAP), 5.6 mg I-17 monomer, and 8.95 mg II-17 monomer were added to 3 mL of 1-ethyl-3-methylimidazolium sulfate ethyl ester ([EMIM][EtSO4]) and stirred until dissolved. 72 μL of an 8 mg / mL lactate oxidase (LOx) aqueous solution was drawn and mixed thoroughly to obtain a mixed solution containing 2.4% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.6 V to +0.9 V, thus obtaining the copolymer enzyme electrode. After LOx was incorporated, the enzyme activity remained above 90%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of LOx remained at 88.9%. At this copolymerization ratio, the QCM chip frequency decreased by 3 Hz, and the charge transfer impedance was 7.07 Ω·cm. -2 The sensing limit for lactic acid is 21 μM, and it retains 82% of its response after 4 weeks.

[0093] Example 21

[0094] In this embodiment, electrochemical copolymerization was employed. 16.92 mg of tetrabutylammonium hexafluorophosphate (TBHAP), 2.15 mg of I-18 monomer, and 10.94 mg of II-18 monomer were added to 3 mL of 1-ethyl-3-methylimidazolium hydrogen sulfate ([EMIM][HSO4]) and stirred until dissolved. 72 μL of a 9 mg / mL laccase aqueous solution was drawn and mixed thoroughly to obtain a mixed solution containing 2.4% water. This 3 mL mixed solution was added to an electrolytic cell. Using a gold electrode as the working electrode, Ag / AgCl as the reference electrode, and a platinum electrode as the counter electrode, the polymerization potential range was -0.7 V to +0.8 V, thus obtaining the copolymer enzyme electrode. After lacase conjugation, the enzyme activity remained above 90%. Ultraviolet absorption spectroscopy after electrochemical polymerization showed that the relative enzyme activity of lacase remained at 89.9%. At this copolymerization ratio, the QCM chip frequency decreased by 5 Hz, and the charge transfer impedance was 6.48 Ω·cm. -2 The sensing limit for adrenaline is 8 μM, and it retains 83% of its response after 4 weeks.

[0095] The monomer structures, enzyme types, and polymerization conditions used in Examples 1-21 are shown in Table 1.

[0096] Table 1. Monomer structural formulas, enzyme types, and polymerization conditions of preferred embodiments of the present invention.

[0097]

[0098]

[0099]

[0100] The embodiments of the present invention have been described above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments. Various changes can be made according to the purpose of the invention. Any changes, modifications, substitutions, combinations or simplifications made based on the spirit and principle of the technical solution of the present invention shall be equivalent substitutions. As long as they meet the purpose of the invention and do not deviate from the technical principle and inventive concept of the present invention, they shall fall within the protection scope of the present invention.

Claims

1. A biomimetic conductive polymer enzyme-based sensing material, characterized in that: This material is a composite material of biomimetic conductive polymer and biological enzyme; wherein the biomimetic conductive polymer contains both zwitterionic groups and redox groups; and contains monomer units as shown in formula (I): in These are groups that form the backbone of a conductive polymer; -L1- is -(CH2) x -、-(CH2) x -O-(CH2) y -、-(CH2) x -O-CO-(CH2) y -、-(CH2) x -(CH2-CH2-O) y -、-(CH2) x -O-(CH2-CH2-O) y -、-O-(CH2) x -(CH2-CH2-O) y -or -O-(CH2) x -O-(CH2-CH2-O) y - any one of the following, where x and y are integers, and 0 ≤ x ≤ 20, 0 ≤ y ≤ 20; -L2- is -(CH2) x -、-O-(CH2) x -、-C(O)-O-(CH2) x -、-C(O)-NH-(CH2) x -、-C(CH3)2-C(O)-O-(CH2) x -、-CH(CH3)-O-(CH2) x -、-S-CH2-CH2-O-(CH2) x -, -S-CH2-CH(CH3)-C(O)-O-(CH2) x - any one or more of the following, where x is an integer and 0 ≤ x ≤ 20; -R1 is a zwitterionic functional group that can resist protein, microbial and cell adhesion; Alternatively, the biomimetic conductive polymer enzyme-based sensing material may contain monomer units as shown in formula (I) as well as monomer units as shown in formula (II): in These are groups that form the backbone of a conductive polymer; -L3- can be any one or more of -CH2-, -O-, -S-, -NH-, -C(O)-, -C(O)NH-, -NH-C(O)-, -OC(O)-, or -C(O)O-; -L4- is -(CH2) x -、-(CH2) x -O-(CH2) y -、-(CH2) x -O-CO-(CH2) y -、-(CH2) x -(CH2-CH2-O) y -、-(CH2) x -O-(CH2-CH2-O) y -、-O-(CH2) x -(CH2-CH2-O) y -or -O-(CH2) x -O-(CH2-CH2-O) y - any one of the following, where x and y are integers, and 0 ≤ x ≤ 20, 0 ≤ y ≤ 20; -R2 is a redox functionalized group; The biomimetic conductive polymer enzyme-based sensing material is prepared by the following method, which involves dissolving one or more monomer units of formula (I) and one or more monomer units of formula (II) in an ionic liquid containing biological enzymes and then performing electrochemical copolymerization deposition to prepare an enzyme-containing copolymer sensing material, including the following steps: (1) Add one or more conductive polymer monomers, biological enzymes, trace amounts of water and electrolytes to the ionic liquid and stir until dissolved; (2) Add the solution obtained in step (1) to the electrolytic cell and adopt a three-electrode system. Set up the working electrode, Ag / AgCl reference electrode and platinum mesh counter electrode. The combined concentration of conductive polymer monomers is 0.1mM to 50mM, the enzyme concentration range is 0.5mg / mL to 15mg / mL, the water content is 0.4% to 4.8%, the electrolyte concentration is 10mM to 100mM, and the polymerization potential range is set to -1V to 3V. Electrochemical polymerization is used to obtain the electrochemical enzyme sensing electrode.

2. The biomimetic conductive polymer enzyme-based sensing material according to claim 1, characterized in that: Its contained monomer units use At least one of them; In its constituent monomer unit structure formula (I), -L1- is -(CH2). x -、-(CH2) x -O-(CH2) y -、-(CH2) x -O-CO-(CH2) y -、-(CH2) x -(CH2-CH2-O) y - any one of the following, where x and y are integers, and 0 ≤ x ≤ 5, 0 ≤ y ≤ 5; In its constituent monomer unit structure formula (I), -L2- is -(CH2). x -、-O-(CH2) x -、-C(O)-O-(CH2) x -、-C(O)-NH-(CH2) x -、-C(CH3)2-C(O)-O-(CH2) x -、-CH(CH3)-O-(CH2) x - any one or more of the following, where x and y are integers, and 0 ≤ x ≤ 5, 0 ≤ y ≤ 5; In its monomer unit structure formula (Ⅱ), -L3- can be any one of -CH2-, -O-, -S-, -NH-, -C(O)-, -NH-C(O)-, and -OC(O)-. In its constituent monomer unit structure formula (II), -L4- is -(CH2). x -、-(CH2) x -O-(CH2) y -、-(CH2) x -O-CO-(CH2) y -、-(CH2) x -(CH2-CH2-O) y -、-O-(CH2) x -(CH2-CH2-O) y - any one of the following, where x and y are integers, and 0 ≤ x ≤ 20, 0 ≤ y ≤ 20.

3. The biomimetic conductive polymer enzyme-based sensing material according to claim 1 or 2, characterized in that: In its constituent monomer unit structure formula (I), -R1 is Any one of them; In its constituent monomer unit structure formula (II), -R2 is Any one of them.

4. A method for preparing the biomimetic conductive polymeric enzyme-based sensing material according to claim 1, characterized in that: The preparation of enzyme-containing copolymer sensing materials involves dissolving one or more monomer units of formula (I) and one or more monomer units of formula (II) in an ionic liquid containing biological enzymes and then performing electrochemical copolymerization deposition, including the following steps: (1) Add one or more conductive polymer monomers, biological enzymes, trace amounts of water and electrolytes to the ionic liquid and stir until dissolved; (2) Add the solution obtained in step (1) to the electrolytic cell and adopt a three-electrode system. Set up the working electrode, Ag / AgCl reference electrode and platinum mesh counter electrode. The combined concentration of conductive polymer monomers is 0.1mM to 50mM, the enzyme concentration range is 0.5mg / mL to 15mg / mL, the water content is 0.4% to 4.8%, the electrolyte concentration is 10mM to 100mM, and the polymerization potential range is set to -1V to 3V. Electrochemical polymerization is used to obtain the electrochemical enzyme sensing electrode.

5. The method for preparing the biomimetic conductive polymer enzyme-based sensing material according to claim 4, characterized in that: The biological enzymes used are at least one of glucose oxidase, glucose dehydrogenase, galactose oxidase, cholesterol oxidase, amino acid oxidase, alcohol oxidase, lactate oxidase, laccase, catalase and uricase. The electrolyte used is at least one of lithium chloride, lithium perchlorate, tetrabutylammonium perchlorate, tetrabutylammonium hexafluorophosphate, tetrabutylammonium tetrafluoroborate, lithium dioxaborate, and lithium bis(trifluoromethanesulfonyl)imide. The ionic liquid is composed of cations and anions and has A + B - The general structural formula, where A + Represents a cation, which is One type; where -R1, -R2, -R3, and -R4 are -H and -(CH2). x -、-(CH3) x -(CH2) x CH3, -C x H 2x+1 N≡C(CH2) x -, -CH=CH-(CH3) x -CH2-CH2-(O-CH2-CH2) x -OH or -CH2-CH2-(O-CH2-CH-(CH3)x) y One or more of -OH groups; where x and y are integers, and 0 ≤ x ≤ 30, 0 ≤ y ≤ 30; B - Represents anion, Br - Cl - I - F - BF4 - PF4 - PF6 - oTf - ,HAc - HSO4 - EtSO4 - SCN - CN - OCN - CNO - CF3SO3 - CF3COO - (CF3SO2)2N - (CF3SO2)2Cl - (HO)2PO2 - AcO - One of them.

6. The method for preparing the biomimetic conductive polymer enzyme-based sensing material according to claim 5, characterized in that: The biological enzyme is at least one of glucose oxidase, lactate oxidase, glucose dehydrogenase, and catalase. The electrolyte is selected based on its solubility and doping effect on electrochemical polymerization. The electrolyte is at least one of lithium perchlorate, tetrabutylammonium perchlorate, and tetrabutylammonium hexafluorophosphate. Ionic liquids are categorized based on their solubility in various monomers, retention of enzyme activity, and controllability of electrochemical polymerization. Ionic liquids utilize at least one of the following six criteria:

7. The method for preparing the biomimetic conductive polymer enzyme-based sensing material according to claim 4, characterized in that: The enzyme concentration used ranges from 1 mg / mL to 10 mg / mL; The polymerization potential range used is -0.8V to 1.4V.

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