CN103263273A - Preparation process of enzyme electrode with biocompatibility - Google Patents

Preparation process of enzyme electrode with biocompatibility Download PDF

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CN103263273A
CN103263273A CN2013102237021A CN201310223702A CN103263273A CN 103263273 A CN103263273 A CN 103263273A CN 2013102237021 A CN2013102237021 A CN 2013102237021A CN 201310223702 A CN201310223702 A CN 201310223702A CN 103263273 A CN103263273 A CN 103263273A
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pmfv
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李子寅
金仲恩
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Suzhou Cosmetic New Materials Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
    • A61B5/1486—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue using enzyme electrodes, e.g. with immobilised oxidase

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Abstract

The invention discloses a preparation process of an enzyme electrode with biocompatibility. The preparation process comprises the following steps of: (1) synthesizing poly-2-methacrylamide oxyethyl phosphorylcholine-vinylferrocene-vinylbenzeneboronic acid (PMFV); (2) synthesizing polyving akohol (PVAN) with active ester groups; (3) preparing cis-polyhydric long-chain glucose oxidase (PVA-GOx); (4) preparing a blood glucose sensor electrode with a PMFV and PVA-GOx alternating arrangement structure, arranging the PMFV layer on the outermost layer of the electrode, and ensuring an MPC unit in the PMFV to form a bionic structure similar to a cytomembrane phospholipid molecular layer on the outermost layer of the electrode, thus effectively inhibiting the adsorption of the protein and cell, preventing the formation of thrombus and inflammation, and improving the biocompatibility of the sensor electrode; and adopting the VFC unit in the PMFV as an electron transmission medium to realize the monitoring of the blood glucose concentration. The thickness of an entire alternating deposition layer is less than 100nm and far smaller than the size of a conventional probe-type electrode.

Description

A kind of preparation technology with enzyme electrode of biocompatibility
Technical field
The invention belongs to medical instruments field, particularly a kind of preparation technology with enzyme electrode of biocompatibility.
Background technology
Biosensor is to utilize biomolecule such as enzyme, antibody as intermediary, utilize specific reactions such as its catalysis or immunity combination with certain composition in the body, concentration information as blood glucose, blood fat or virus etc. changes current signal into, thereby realizes the purpose to the real-time monitoring of a certain biochemical index in the body.The most representative in the biosensor is blood glucose sensor, and traditional enzyme electrode blood glucose sensor uses the metal electrode that has glucoseoxidase (GOx) coating as the blood glucose measurement electrode, and its shortcoming has the following aspects: directly use O 1. 2/ H 2O 2Transmit intermediary as electronics, measuring process needs external voltage higher, and measurement result is subjected to the influence that oxygen concentration changes in the blood easily, and measurement result is not accurate enough.2. glucoseoxidase (GOx) needs to be coated and fixed by semipermeable membrane, and this semipermeable membrane can cause serum albumin in the absorption on surface, makes micromolecularly to cause the transducer sensitivity variation through being obstructed, respond slack-off, adverse consequencess such as signal to noise ratio reduction.3. the electrode biocompatibility is poor, because the immunologic rejection effect causes untoward reaction such as thrombosis and inflammation easily.Thereby, seek suitable electronics and transmit intermediary, using stable enzyme fixing means and the biocompatibility that improves sensor electrode is the focal point of present biosensor development work.
Mentioned in some inventions and utilized the electric conductivity hydrogel that glucoseoxidase (GOx) is coated on the method for electrode surface, the electronics in the middle of the hydrogel transmits intermediary such as ferrocene has replaced O 2/ H 2O 2, hydrogel network allows in the body fluid micromolecular penetrating in immobilized enzyme, and these schemes have solved above-mentioned problem 1 and problem 2, but the biocompatibility of this class blood glucose sensor still awaits further improvement.
Summary of the invention
At deficiency of the prior art, the invention provides a kind of preparation technology with enzyme electrode of biocompatibility, this technology can be improved fixing, micromolecular permeability and the biocompatibility of enzyme simultaneously.
This preparation technology's concrete steps are as follows:
Step 1: synthetic poly-2-methylacryoyloxyethyl phosphocholine-vinyl ferrocene-vinylphenylboronic acid (PMFV):
2-methylacryoyloxyethyl phosphocholine (MPC), vinyl ferrocene (VFC) and vinylphenylboronic acid (VPBA) are mixed with alcoholic solution in proportion, in alcoholic solution, add polymerization initiator, in having the environment of protective gas, carry out polyreaction, with the polymerizate precipitate and separate, and carry out vacuum drying;
Step 2: synthetic a kind of polyvinyl alcohol (PVAN) that has the active ester group:
Polyvinyl alcohol is dissolved in anhydrous adjacent methyl pyrrolidone (NMP), and solution slowly adds anhydrous pyridine and nitrophenol chloromethyl ester (NPC) in proportion when stirring, and after a period of time is carried out in reaction, isolates polyvinyl alcohol (PVAN) with ether sedimentation;
Step 3: preparation has the glucoseoxidase (PVA-GOx) of many hydroxyls of cis long-chain:
Glucoseoxidase (GOx) is joined in the phosphate buffer solution (PBS) of pH=8-10, add polyvinyl alcohol (PVAN) in proportion, stir a period of time, amido reaction in active ester unit in the polyvinyl alcohol (PVAN) and the glucoseoxidase (GOx), the reaction back uses molecular cut off to remove unreacted polyvinyl alcohol (PVAN) and other by-product in the dialyzer dialysis of 50K-150K, obtains glucoseoxidase (PVA-GOx) solid by lyophilization;
Step 4: the glucoseoxidase (PVA-GOx) that preparation has poly-2-methylacryoyloxyethyl phosphocholine-vinyl ferrocene-vinylphenylboronic acid (PMFV) and has many hydroxyls of cis long-chain replaces the blood glucose sensor electrode of arrangement architecture:
Poly-2-methylacryoyloxyethyl phosphocholine-vinyl ferrocene-vinylphenylboronic acid (PMFV) and the glucoseoxidase (PVA-GOx) that has many hydroxyls of cis long-chain successively are deposited on gold electrode surfaces by the covalent bond of vinylphenylboronic acid (VPBA) and the many hydroxyls of cis, and outermost layer is for gathering 2-methylacryoyloxyethyl phosphocholine-vinyl ferrocene-vinylphenylboronic acid (PMFV) layer.
Preferably; in step 1, described polyreaction, is carried out under 65 ℃ and is protective gas with the argon as initiator with azo-compound; the polymerizate that polyreaction obtains further carries out precipitate and separate with the EC miscible agent, and vacuum drying 24 hours.
Preferably, in step 2, solution slowly adds anhydrous pyridine and nitrophenol chloromethyl ester (NPC) in proportion in 0 ℃ of stirring, is reflected at and carries out under 0 ℃ 5 hours, and isolate polyvinyl alcohol (PVAN) with ether sedimentation.
Preferably, in step 3, glucoseoxidase (GOx) is joined in the phosphate buffer solution (PBS) of pH=8.4, add polyvinyl alcohol (PVAN) in proportion, exist again
4 ℃ were stirred 12 hours down, amido reaction in active ester unit in the polyvinyl alcohol (PVAN) and the glucoseoxidase (GOx), the reaction back uses molecular cut off to remove unreacted polyvinyl alcohol (PVAN) and other by-product as the dialyzer dialysis of 100K, obtains having glucoseoxidase (PVA-GOx) solid of many hydroxyls of cis long-chain by lyophilization.
Preferably, in step 4, poly-2-methylacryoyloxyethyl phosphocholine-vinyl ferrocene-vinylphenylboronic acid (PMFV) and the arrangement architecture that replaces that has a glucoseoxidase (PVA-GOx) of many hydroxyls of cis long-chain are made by sedimentation (Layer-by-Layer Deposition) successively.
The thickness limits of the alternating deposition layer on the blood glucose sensor electrode that makes with above-mentioned technology is in 100nm, and technique effect is preferable.
Compared with prior art, advantage of the present invention is:
By preparation technology of the present invention, the glucoseoxidase (PVA-GOx) that preparation has poly-2-methylacryoyloxyethyl phosphocholine-vinyl ferrocene-vinylphenylboronic acid (PMFV) and has many hydroxyls of cis long-chain replaces the blood glucose sensor electrode of arrangement architecture, wherein, MFV and PVA-GOx can successively be deposited on gold electrode surfaces by the covalent bond of vinylphenylboronic acid (VPBA) and the many hydroxyls of cis, and to require outermost layer be the PMFV layer.MPC unit among the PMFV can form the biomimetic features that is similar to the cell membrane phospholipid molecular layer at the electrode outermost layer, the effectively absorption of Profilin and cell, the formation of anti-tampon and inflammation, the biocompatibility of raising sensor electrode.GOx is fixed in the polymeric layer network by the combination with PVA, and this network structure can allow small-molecule substance such as glucose, the freeing in and out of oxygen etc.VFC unit among the PMFV transmits intermediary as electronics, has constituted the electronics transmission path of glucose → GOx → VFC → gold electrode surfaces, has realized the monitoring to blood sugar concentration.The thickness of whole polymeric layer is in 100nm, much smaller than the size of common probe-type electrode.
Description of drawings
Fig. 1 is the sketch map of PVAN and GOx reaction mechanism.
Fig. 2 is the enzyme electrode structural representation of the present invention's preparation.
Fig. 3 is the result schematic diagram of the concentration of glucose test experiments of experimental electrode in serum albumin solution.
Fig. 4 is the matched curve of electric current density and concentration of glucose inverse.
Fig. 5 is the handling process sketch map of spun gold electrode.
Fig. 6 is the handling process sketch map of gold-plated electrode of substrate.
The specific embodiment
A kind of biological sensor electrode preparation method of the present invention, concrete steps are as follows:
1. synthesize poly-2-methylacryoyloxyethyl phosphocholine-vinyl ferrocene-vinylphenylboronic acid (PMFV): 2-methylacryoyloxyethyl phosphocholine (MPC), vinyl ferrocene (VFC) and vinylphenylboronic acid (VPBA) are mixed with alcoholic solution in proportion; in solution, add azodiisobutyronitrile (AIBN) as polymerization initiator; polyreaction is carried out under 65 ℃ and is protective gas with the argon; polymerizate carries out precipitate and separate with the EC miscible agent, and vacuum drying 24 hours.Wherein, the structure of PMFV is:
Figure BSA00000907761000051
2. synthesize a kind of polyvinyl alcohol (PVAN) that has active ester group: polyvinyl alcohol is dissolved in anhydrous adjacent methyl pyrrolidone (NMP), solution slowly adds anhydrous pyridine and nitrophenol chloromethyl ester (NPC) in proportion in 0 ℃ of stirring, be reflected at and carried out under 0 ℃ 5 hours, isolate polyvinyl alcohol (PVAN) with ether sedimentation.Wherein, the structure of polyvinyl alcohol (PVAN) is:
Figure BSA00000907761000061
3. preparation has the glucoseoxidase PVA-GOx of many hydroxyls of cis long-chain: GOx is joined in the phosphate buffer solution (PBS) of pH=8.4, add PVAN in proportion, stirred 12 hours at 4 ℃, mechanism and the reaction of the amido among the GOx in the accompanying drawing 1 pressed in active ester unit among the PVAN, the reaction back uses molecular cut off to remove unreacted PVA and other by-product as the dialyzer dialysis of 100K, obtains the PVA-GOx solid by lyophilization.
4. preparation has the blood glucose sensor electrode that PMFV and PVA-GOx replace arrangement architecture: as shown in Figure 2, PMFV and PVA-GOx can successively be deposited on gold electrode surfaces by the covalent bond of vinylphenylboronic acid (VPBA) and the many hydroxyls of cis, and to require outermost layer be the PMFV layer.MPC unit among the PMFV can form the biomimetic features that is similar to the cell membrane phospholipid molecular layer at the electrode outermost layer, the effectively absorption of Profilin and cell, the formation of anti-tampon and inflammation, the biocompatibility of raising sensor electrode.GOx is fixed in the polymeric layer network by the combination with PVA, and this network structure can allow small-molecule substance such as glucose, the freeing in and out of oxygen etc.VFC unit among the PMFV transmits intermediary as electronics, has constituted the electronics transmission path of glucose → GOx → VFC → gold electrode surfaces, has realized the monitoring to blood sugar concentration.The thickness of whole alternating deposition layer is in 100nm, much smaller than the size of common probe-type electrode.
Its technique effect is analyzed as follows:
1. the blood glucose sensor electrode of making is placed serum albumin solution, concentration by 1mmol/L in continuing the process that stirs adds glucose one by one, from accompanying drawing 3 as can be seen, the electric current amplitude of variation that changes corresponding to concentration of glucose is higher than 10 times of current noise amplitude, and response time was less than 20 seconds, show this electrode reaction sensitivity, the signal to noise ratio height.
2. for desirable enzyme electrode, its electric current density should satisfy Michaelis-Menten equation (Michaelis-Menten Equation) with the concentration relationship of measuring substrate:
Figure BSA00000907761000071
Wherein J is electric current density, and [S] for measuring concentration of substrate, Km and Jmax are constant.The size of current of correspondence during measure glucose concentration 2mmol/L to 12mmol/L under above-mentioned experiment condition, and get its inverse and carry out linear fit, fitting result as shown in Figure 4, R=0.998 shows that the measurement result accuracy of this electrode is good.
3. the gold-plated section that the surface is had the gold-plated section of above-mentioned polymer coating and do not do any processing was immersed down in the blood plasma that contains anticoagulant 30 minutes at 37 ℃.As can be seen the slice surface platelet adsorbance less (data result sees Table 1) of polymer coating is being arranged by stereoscan photograph, and do not occurring significantly activating and assembling, and the absorption of undressed slice surface platelet is more,
And tangible pseudopodium and gathering appearred.As seen this invention can significantly improve the biocompatibility of biological sensor electrode.
The platelet adsorbance of table 1 polymer electrode and gold-plated substrate surface
Figure BSA00000907761000081
At this, provide following several preparation method:
Embodiment 1
1. with 2.95gMPC, 1.27gVFC, 0.58gVPBA and 16mgAIBN is dissolved in the 20mL dehydrated alcohol, solution is transferred in the 50mL teat glass, continue in the test tube to feed argon after 3 minutes with the test tube sealing, test tube left standstill after 72 hours 65 ℃ of oil baths and to be cooled to room temperature, with 450mL ether+150mL chloroform mixed liquor as precipitant, reaction solution is dropwise joined in the precipitant, with no slag filter paper filtering and collecting precipitation, will precipitate vacuum drying and obtain the PMFV solid.
2. be that 1700 PVA is dissolved among 100mL80 ℃ the anhydrous NMP with the 2.00g degree of polymerization, treat to add the 0.27g anhydrous pyridine after solution left standstill is cooled to room temperature, solution when stirring, ice bath is slowly added 0.71gNPC, react to react after 5 hours to dissolve and dropwise join in the 800mL absolute ether, with no slag filter paper filtering and collecting precipitation, obtain the PVAN solid after being deposited in vacuum drying.
3. 20mgGOx is dissolved among the PBS that 2mL concentration is 0.1mol/L pH=8.4, add 30mgPVAN, solution was stirred 12 hours at 4 ℃, with deionized water solution dilution is arrived 5mL, use molecular cut off to dialyse 72 hours as the bag filter of 100K, the solution lyophilization in the bag filter is obtained the PVA-GOx solid.
4. the 0.5mm spun gold electrode after will polishing is to soak 8 hours in the alcoholic solution of 20mmol/L in 3-mercaptopropionic acid ethyl ester concentration, cleaning the back with ethanol soaked 5 minutes in mass fraction is 0.2% photoreactivity PVA (a kind of water-soluble poval with azido group) aqueous solution, using wavelength 250nm, power after taking out from solution is 80mW/cm 2 Ultraviolet source irradiation 1 minute, and with distilled water flushing 3 minutes.
5. PMFV and PVA-GOx are mixed with the aqueous solution that mass concentration is 2.5mg/mL respectively.Finish the preparation of electrode by the route in the accompanying drawing 6, namely, spun gold electrode after will handling by this embodiment, after mass concentration was to soak 1 hour in the PMFV solution of 2.5mg/mL, with distilled water immersion 10 minutes, the GOx solution of putting into 2.5mg/mL again soaked after 1 hour, with distilled water immersion 10 minutes, then get back to and put into the link that PMFV solution soaks, so repeat 8-12 circulation after, namely making required is the enzyme electrode of matrix with the spun gold electrode.
Embodiment 2
0.5mm spun gold electrode after the polishing is to soak 8 hours in the alcoholic solution of 20mmol/L in 2-aminoothyl mercaptan concentration, cleaning the back with ethanol soaked 5 minutes in the PVAN mass fraction is 0.25% 2-methylformamide solution, take out dry back and in concentration is the sodium hydrate aqueous solution of 1mmol/L, soaked 10 minutes, and with distilled water flushing 3 minutes.Press the synthetic PMFV of method and PVA-GOx among the embodiment 1, and it is mixed with mass fraction respectively is 0.25% aqueous solution.Finish the preparation of electrode by the route in the accompanying drawing 6, namely, spun gold electrode after will handling by this embodiment, after in mass fraction is the PMFV solution of 0.25% 2.5mg/mL, soaking 1 hour, with distilled water immersion 10 minutes, the GOx solution of putting into mass fraction again and be 0.25% 2.5mg/mL soaked after 1 hour, with distilled water immersion 10 minutes, then return and put into the link that PMFV solution soaks, so repeat 8-12 circulate after, making required is the enzyme electrode of matrix with the spun gold electrode.
Embodiment 3
Gold-plated substrate is to soak 8 hours in the alcoholic solution of 20mmol/L in 3-mercaptopropionic acid ethyl ester concentration, cleaning the back with ethanol covers to the surface fully at the photoreactivity PVA of surface dropping 0.2%, use wavelength 250nm, power is the ultraviolet source irradiation 1 minute of 80mW/cm2, and with distilled water flushing 3 minutes.By the synthetic PMFV of the method in the example one and PVA-GOx, and it is mixed with mass fraction respectively is 0.25% aqueous solution.Finish the preparation of electrode then by the route in the accompanying drawing 6, namely, on the gold-plated substrate after this embodiment is handled, the dropping mass fraction is 0.25% PMFV solution, behind the speed rotary plating with 1 minute 500rpm+30 3000rpm second, drip distilled water and left standstill 3 minutes at substrate surface, the speed with 30 seconds 3000rpm dries again; Then drip mass fraction and be 0.25% GOx solution, behind the speed rotary plating with 1 minute 500rpm+30 3000rpm second, the dropping distilled water also left standstill 3 minutes, speed with 30 seconds 3000rpm dries again, return the link that drips PMFV solution, so repeat 8-12 circulate after, namely making required is the enzyme electrode of matrix with gold-plated substrate.
Embodiment 4
Gold-plated substrate is to soak 8 hours in the alcoholic solution of 20mmol/L in 2-aminoothyl mercaptan concentration, cleaning the back with ethanol, drip polyvinyl alcohol (PVAN) mass fraction on the surface be that 0.25% 2-methylformamide solution covers fully to the surface, use rotary plating method (1 minute 500rpm+30 3000rpm second) to form polyvinyl alcohol (PVAN) thin film on the surface, be to soak 10 minutes in the sodium hydrate aqueous solution of 1mmol/L in concentration, and with distilled water flushing 3 minutes.By the synthetic PMFV of the method in the example one and PVA-GOx, and it is mixed with mass fraction respectively is 0.25% aqueous solution.Finish the preparation of electrode then by the route in the accompanying drawing 6, namely, on the gold-plated substrate after this embodiment is handled, the dropping mass fraction is 0.25% PMFV solution, behind the speed rotary plating with 1 minute 500rpm+30 3000rpm second, drip distilled water and left standstill 3 minutes at substrate surface, the speed with 30 seconds 3000rpm dries again; Then drip mass fraction and be 0.25% GOx solution, behind the speed rotary plating with 1 minute 500rpm+30 3000rpm second, the dropping distilled water also left standstill 3 minutes, speed with 30 seconds 3000rpm dries again, return the link that drips PMFV solution, so repeat 8-12 circulate after, namely making required is the enzyme electrode of matrix with gold-plated substrate.
The above; it only is preferable case study on implementation of the present invention; be not that the present invention is imposed any restrictions; every similar mode of any simple modification, change, employing that essence is done above embodiment according to the present invention substitutes and the variation of equivalent structure, all still belongs in the protection domain of technical solution of the present invention.

Claims (8)

1. preparation technology with enzyme electrode of biocompatibility is characterized in that concrete steps are as follows:
Step 1: synthetic poly-2-methylacryoyloxyethyl phosphocholine-vinyl ferrocene-vinylphenylboronic acid (PMFV):
2-methylacryoyloxyethyl phosphocholine (MPC), vinyl ferrocene (VFC) and vinylphenylboronic acid (VPBA) are mixed with alcoholic solution in proportion, in alcoholic solution, add polymerization initiator, in having the environment of protective gas, carry out polyreaction, with the polymerizate precipitate and separate, and carry out vacuum drying;
Step 2: synthetic a kind of polyvinyl alcohol (PVAN) that has the active ester group:
Polyvinyl alcohol is dissolved in anhydrous adjacent methyl pyrrolidone (NMP), and solution slowly adds anhydrous pyridine and nitrophenol chloromethyl ester (NPC) in proportion when stirring, and after a period of time is carried out in reaction, isolates polyvinyl alcohol (PVAN) with ether sedimentation;
Step 3: preparation has the glucoseoxidase (PVA-GOx) of many hydroxyls of cis long-chain:
Glucoseoxidase (GOx) is joined in the phosphate buffer solution (PBS) of pH=8-10, add polyvinyl alcohol (PVAN) in proportion, stir a period of time, amido reaction in active ester unit in the polyvinyl alcohol (PVAN) and the glucoseoxidase (GOx), the reaction back uses molecular cut off to remove unreacted polyvinyl alcohol (PVAN) and other by-product in the dialyzer dialysis of 50K-150K, obtains glucoseoxidase (PVA-GOx) solid by lyophilization;
Step 4: the glucoseoxidase (PVA-GOx) that preparation has poly-2-methylacryoyloxyethyl phosphocholine-vinyl ferrocene-vinylphenylboronic acid (PMFV) and has many hydroxyls of cis long-chain replaces the blood glucose sensor electrode of arrangement architecture:
Poly-2-methylacryoyloxyethyl phosphocholine-vinyl ferrocene-vinylphenylboronic acid (PMFV) and the glucoseoxidase (PVA-GOx) that has many hydroxyls of cis long-chain successively are deposited on gold electrode surfaces by the covalent bond of vinylphenylboronic acid (VPBA) and the many hydroxyls of cis, and outermost layer is for gathering 2-methylacryoyloxyethyl phosphocholine-vinyl ferrocene-vinylphenylboronic acid (PMFV) layer.
2. the preparation technology with enzyme electrode of biocompatibility according to claim 1 is characterized in that, in the step 1, described polyreaction, is carried out under 65 ℃ and is protective gas with the argon as initiator with azo compound.
3. the preparation technology with enzyme electrode of biocompatibility according to claim 1 and 2 is characterized in that, in the step 1, the polymerizate that polyreaction is obtained carries out precipitate and separate with the EC miscible agent, and vacuum drying 24 hours.
4. the preparation technology with enzyme electrode of biocompatibility according to claim 1, it is characterized in that, in the step 2, solution slowly adds anhydrous pyridine and nitrophenol chloromethyl ester (NPC) in proportion in 0 ℃ of stirring, be reflected at and carried out under 0 ℃ 5 hours, and isolate polyvinyl alcohol (PVAN) with ether sedimentation.
5. the preparation technology with enzyme electrode of biocompatibility according to claim 1 is characterized in that, in the step 3, glucoseoxidase (GOx) is joined in the phosphate buffer solution (PBS) of pH=8.4, adds polyvinyl alcohol (PVAN) in proportion.
6. the preparation technology with enzyme electrode of biocompatibility according to claim 5, it is characterized in that, in the step 3, after adding polyvinyl alcohol (PVAN) in proportion, stirred 12 hours down at 4 ℃ again, amido reaction in active ester unit in the polyvinyl alcohol (PVAN) and the glucoseoxidase (GOx), the reaction back uses molecular cut off to remove unreacted polyvinyl alcohol (PVAN) and other by-product as the dialyzer dialysis of 100K, obtains having glucoseoxidase (PVA-GOx) solid of many hydroxyls of cis long-chain by lyophilization.
7. the preparation technology with enzyme electrode of biocompatibility according to claim 1, it is characterized in that, in the step 4, poly-2-methylacryoyloxyethyl phosphocholine-vinyl ferrocene-vinylphenylboronic acid (PMFV) and the alternately arrangement architecture that has a glucoseoxidase (PVA-GOx) of many hydroxyls of cis long-chain prepare by sedimentation successively.
8. the preparation technology with enzyme electrode of biocompatibility according to claim 1 is characterized in that, replaces the thickness of illuvium on the blood glucose sensor electrode in 100nm.
CN2013102237021A 2013-06-07 2013-06-07 Preparation process of enzyme electrode with biocompatibility Pending CN103263273A (en)

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CN107296614A (en) * 2017-05-22 2017-10-27 中山大学 Blood sugar monitor based on biocompatibility electrode
CN109860592A (en) * 2018-12-25 2019-06-07 上海力信能源科技有限责任公司 A kind of nickel-cobalt lithium manganate cathode material and preparation method thereof of boracic molecular modification

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Publication number Priority date Publication date Assignee Title
CN105680056A (en) * 2016-01-19 2016-06-15 苏州蔻美新材料有限公司 Preparation method for anode device of microbial fuel cell
CN107296614A (en) * 2017-05-22 2017-10-27 中山大学 Blood sugar monitor based on biocompatibility electrode
CN109860592A (en) * 2018-12-25 2019-06-07 上海力信能源科技有限责任公司 A kind of nickel-cobalt lithium manganate cathode material and preparation method thereof of boracic molecular modification

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Application publication date: 20130828