CN106525788A - Preparation method of bionic nanometer film and method for fixing probe with same - Google Patents
Preparation method of bionic nanometer film and method for fixing probe with same Download PDFInfo
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Abstract
The invention discloses a preparation method of a bionic nanometer film and a method for fixing probes with the bionic nanometer film. The purpose is to solve the problems that according to an existing modifying method, the environment is severely polluted, and the modifying process is complex. First, a polydopamine film is formed on the surface of a biochip, and the biomolecule probes are fixed to the active surface of the biochip. According to the construction method, the polydopamine film can be formed on the surfaces of kinds of organic and inorganic materials without carrying out complex pretreatment on the surface of a substrate, reaction conditions are mild, it is avoided that organic solvent for usage pollutes the environment, and therefore the method has the advantages of being economical, efficient, bionic, environmentally friendly and easy and convenient to operate. The surface of polydopamine has rich active functional groups, the load capacity of the biomolecule probes can be increased, the fixing efficiency is improved for the biomolecule probes, and the biological activity of the biomolecule probes can be effectively kept. Compared with a traditional preparation method, the method makes fixed biomolecules more stable and higher in activity and practicability.
Description
Technical field
The invention belongs to material and field of biosensors, more particularly to a kind of preparation method of bionic nano thin film
And the method for fixing probe with the thin film.
Background technology
Biochip is a kind of trace analysis being with historically new significance, and obtains extensively should in increasing field
With the first step of biochip development is biochip carrier to be prepared, and in the process, needs to carry out chemistry to carrier surface
Process, make it special, fast and efficiently fix various probes.The quality of carrier directly affects the performance of microarray, to chip
Prepare most important, it directly affects the bond strength and efficiency of probe and microscope slide, and then affects the sensitive of results of hybridization
Degree and accuracy.At present, carrier surface processing method mainly has physical absorption and chemical coupling（It is covalently attached）Two big class, tradition
Physical adsorption process it is simple to operate, but have that link efficiency is low, stability is poor, the shortcomings of be susceptible to non-specific adsorption.
Chemical conjugation methods（I.e. in carrier surface growth activity group, the active group of carrier surface is made to pass through covalent cross-linking effectively
Fixed probe）, the method stable connection, but complex steps, the chemical reagent toxicity of employing is higher, sterically hindered than larger,
Modification of surfaces changes native conformation of biomolecule etc..At present, chip surface chemistry is both for a certain specific biological point
Son, but biomolecule of different nature is assembled simultaneously not yet has document report with identical carrier surface, therefore develop a kind of
Environmental protection, the surface modification of easy to operate, highly versatile and fixing meanss become one of focus of research.
In recent years, with the development and the continuous development of nano material preparation technology of nano science, nano material is only with which
Special physicochemical characteristicss and be widely used in the surface modification of biochip, people are devoted to finding with good always
The new method of modifying of biocompatibility.
Existing method of modifying is higher to the profile and dimensional requirement of material, in addition it is also necessary to which complicated pretreatment, what is used has
Machine solvent causes severe contamination to environment, and modification is complicated, and once modification can fix a kind of probe molecule.
The content of the invention
It is an object of the present invention to provide a kind of simple to operate, economical and efficient, the preparation side of the bionic nano thin film of environmental protection
Method, to solve the problems, such as that what existing method of modifying was present causes severe contamination, modification complicated to environment.
It is a further object to provide a kind of method for fixing probe with the thin film.
Technical solution of the present invention is as follows：A kind of preparation method of bionic nano thin film, comprises the steps of：
The pretreatment of A, carrier：Ultrapure water is used after optics microscope slide is soaked with ammonia washing liquid, then is immersed in hydrochloric acid solution
In, then with ultrapure water, N2Dry up；
B, dopamine-hydrochloric acid solution is prepared：Dopamine-hydrochloric acid is weighed, is configured in being dissolved completely in Tirs-HCl buffer
Dopamine-the hydrochloric acid solution of 0.5-4 mg/ milliliters；
C, will be added in dopamine-hydrochloric acid solution through the microscope slide of pretreatment, shake in 50-70 DEG C of thermostatic control oscillator vibration
After shaking 1-48h, rinsed with high purity water, N2Dry up.
Preferably, in step A, ammonia washing liquid is made up of the ammonia and hydrogen peroxide and water that volume ratio is 1: 1: 5.
Preferably, in step A, hydrochloric acid solution is made up of the hydrochloric acid, hydrogen peroxide and water that volume ratio is 1: 1: 5.
Preferably, in step B, dopamine-hydrochloric acid solution process for preparation, to weigh dopamine-hydrochloric acid, is dissolved completely in
PH value be 8.5 Tirs-HCl buffer in be configured to the dopamine-hydrochloric acid solution of 2mg/ milliliters.
Preferably, the time is shaken in step C for 24h.
Preferably, in step C, the temperature of water bath with thermostatic control is 60 DEG C, and the speed of thermostatic control oscillator vibration is 70 rpm.
The method that probe is fixed with thin film obtained above, probe are Fluorescein isothiocyanate 5,20 terminal modified amino
The single stranded DNA of length, is linked on the microscope slide of poly-dopamine modification, and concrete grammar is as follows：
The preparation of A, carbonate buffer solution：0.6g natrium carbonicum calcinatums and 3.7g sodium bicarbonate are dissolved in into 80 milliliters of deionizations respectively
After in water, constant volume to 100 milliliters；
B, fluorescent probe solution are prepared：Take during 10 jig probes are dissolved in 1 milliliter of carbonate buffer solution and be made into 10 mcg/mls
Solution；
The link of C, probe：The slide that poly-dopamine is modified is dipped in fluorescent probe solution, is shaken in the shaking table of 70 rpm
After 1-8h, deionized water is rinsed, and washes unconjugated fluorescent probe off.
Preferably, the method that thin film fixes probe, in step C, the shaking table shaking time is 4h.
Reaction condition of the present invention is gentle, it is to avoid use organic solvent pollution on the environment, and to material surface
Modified to settle at one go, operating procedure is simple, and reaction condition is easily controllable with modifying process.
It is prepared by the main principle for auto polymerization formation poly-dopamine thin film occurring by dopamine in alkaline solution of the invention,
The dihydroxyphenylalanine of the process simulation marine mussel secretion（DOPA）, the principle for adhering in the seawater, the material can make
Mussel strong adsorption is in various body surfaces, and the profile and size to material is not limited, and whole process is complete in aqueous
Into;Simultaneously its powerful adhesive capacity can be modified to all of organic and inorganic materials surface, with versatility and general
And property.Therefore economical and efficient, environmental protection, easy to operate, by assay optimization preparation condition, can be while being adapted to so as to develop
The new method of fixed heterogeneity biomolecule.
Poly-dopamine almost can form the compound of one layer of superpower attachment in any material surface and receive as biomimetic material
Rice thin film, and the profile and size to material do not limit, the application is first by the bionical auto polymerization of this high bioaffinity
Material is introduced in biochip surface modification, to the immobilization amount, activity and the stability that improve biomolecule.
The bionical poly-dopamine nano thin-film of the present invention modifies chip surface, by from poly- complex technique in chip surface shape
Into bionic nano thin film, with the active group fixing biological molecules of film surface.Contain abundant in poly-dopamine nano thin-film surface
Active group, in aqueous solution, there is oxidation polymerization-cross-linking reaction in dopamine, contain in the compound poly-dopamine layer of material surface
Have abundant catechol group, under alkaline environment, these groups are easily oxidized into quinoid structure, so as to contain sulfur
Alcohol (- SH), amino (- NH2), or DNA probe generation Michael's addition (Michael addition) of imino group (- NH)
With schiff base reaction (Schiff base reaction), material surface is introduced biomolecules into, various biomolecules can be met
Fixed requirement, immobilization demand of the simple realization to chip surface biomolecule.
The nano thin-film that the present invention is formed, is mainly made up of 15-20nm nano-particle, and the structure has bigger specific surface
Product, can be with reference to more fluorescent probes.Meanwhile, the membrane structure of granule composition has lower sterically hindered, raising fluorescence
The fixed efficiency and fixed amount of probe, therefore the method is better than other fixing meanss.
In the preparation of above-mentioned bionic nano thin film, optimum condition is：The optium concentration of dopamine-hydrochloric acid solution is 2 mg/ millis
Rise, pH is 8.5, soak time is 24 h（I.e. soak time is 24h）.I.e. the condition of embodiment 15, forms through above-mentioned condition
Nano thin-film, the nano thin-film after polymerization is characterized by SEM, find as shown in figure 1, A, B are respectively sample in Fig. 1
Amplify 10000 times and amplify 50000 times of picture, from Figure 1A as can be seen that sample nano thin-film surface by it is uniform, fine and close
Granule constitute.When being amplified to 50000 times, it can be seen that particle size is homogeneous, grain diameter size is 15-20 nm, is had no
Agglomeration（Figure 1B）.Test result indicate that by the way of the bionic can the surface of matrix formed one layer it is uniform,
Fine and close thin film, is that good platform has been built in the fixation of follow-up fluorescent probe.
The chemical bond composition of slide is modified by infrared spectrum analysiss poly-dopamine further, Fig. 2 is dopamine and polymerization DOPA
The infrared spectrogram of amine.As seen from the figure, there is peak in 3437cm-1 in dopamine, is hydroxyl and amino stretching vibration peak.And be polymerized
There is a spike at 3343 cm-1 in dopamine, is amino stretching vibration peak, may be more to contain in polymer after polymerization
Amino.There is a peak at 1609 cm-1 simultaneously in the two, is the flexural vibrations peak of NH2.Compared with dopamine, gather many
There is a new peak at 1504cm-1 in bar amine, is phenyl ring vibration peak.Result above shows that dopamine successful polymerization is poly- DOPA
Amine.
The optimization of the preparation condition of poly-dopamine nano thin-film, is mainly used to study its fixed effect to probe.First
With fixing meanss conventional at present（Amido modified method and polylysine modification method）Comparison diagram see Fig. 3, from fluorescence microscope
Optical photograph on contrast find, through poly-dopamine modification surface of glass slide than unmodified slide, amido modified slide
It is and there is finer and close fluorescence bright spot in the surface of glass slide of polylysine modification, can fix more fluorescent probes and glimmering
The fluorescence intensity of light probe is high, and phosphor dot is more evenly distributed.Using fluorescence bright spot number of the Image J softwares to above-mentioned four kinds of modes
Amount carries out statistic analysis result such as table 1, and table 1 illustrates that method of modifying of the present invention is contrasted with traditional method of modifying, highlights poly-
The advantage that dopamine is modified in chip surface.
Description of the drawings
Fig. 1 is the scanning electron microscope (SEM) photograph of the nano thin-film of embodiment 15；
Fig. 2 is the infrared spectrogram of dopamine and polymerization dopamine；
Fig. 3 is comparison diagram of the method with common fixation methods that embodiment 15 fixes probe；
Fig. 4 is fluorescent microscopy images figure of the different dopamine concentrations to the impact effect of fixed probe in embodiment 1-4；
Fig. 5 is fluorescent microscopy images figure of the different polymerization times to the impact effect of fixed probe in embodiment 5-10；
Fig. 6 is fluorescent microscopy images figure of the different set times to the impact effect of fixed probe in embodiment 11-14.
Specific embodiment
The following examples can further illustrate the present invention, but limit the present invention never in any form.
Specific embodiment is divided into two steps, and preparation of the first step for bionic nano thin film, second step are the fixation of probe molecule,
The purpose of chip surface modification is exactly that the effect of the preparation of nano thin-film must be by fixed spy in order to fix various molecular probes
Fluorescence after pin counts effect to react.
Embodiment 1,
1st, chip surface is modified with the preparation of bionic nano thin film：
The pretreatment of A, carrier；25mm × 25mm optics microscope slide ammonia washing liquids (volume ratio is ammonia: hydrogen peroxide: water=1:
1: use ultrapure water after 5) soaking 1h, then (volume ratio is hydrochloric acid: hydrogen peroxide: water=1: 1: 5) to be immersed in hydrochloric acid solution
Ultrapure water, N are used after middle 1h2Dry up；
B, dopamine-hydrochloric acid solution is prepared：0.5mg dopamine-hydrochloric acid is accurately weighed, the Tirs-HCl of pH8.5 is dissolved completely in
Dopamine-the hydrochloric acid solution of 0.5 mg/ milliliters is prepared in buffer；
C, will be added in dopamine-hydrochloric acid solution through the microscope slide of pretreatment, temperature（60℃）, thermostatic control oscillator vibration 70
After rpm shakes 24 h, rinsed to face without unnecessary poly-dopamine, N with high purity water2Dry up.
2nd, the fixing meanss of fluorescent probe：Probe is Fluorescein isothiocyanate（FITC）5,20 terminal modified amino length
DNA, be linked to poly-dopamine modification slide on concrete grammar it is as follows,
The preparation of 9.0 carbonate buffer solution of A, 0.5M, pH：0.6g natrium carbonicum calcinatums and 3.7g sodium bicarbonate are dissolved in into 80 respectively
After in ml deionized water, constant volume to 100 milliliters.
B, fluorescent probe solution are prepared：Take during 10 microgram fluorescent probes are dissolved in 1 milliliter of carbonate buffer solution that to be made into 10 micro-
The solution of grams per milliliter.
The link of C, fluorescent probe：The slide that poly-dopamine is modified is dipped in fluorescent probe solution, shaking in 70rpm
After bed shaking 4h, deionized water is repeatedly rinsed, and washes unconjugated fluorescent probe off.
Embodiment 2-4
Embodiment 2-4 step only changes the concentration of the dopamine-hydrochloric acid solution in step B, inquires into optimal with embodiment 1
Dopamine consumption, fluorescence imaging such as Fig. 4 are counted to the fluorescence bright spot quantity of above-mentioned four kinds of modes using Image J softwares
Analysis, as a result such as table 2.When the concentration of dopamine is 0.5mg/ milliliters, phosphor dot negligible amounts, and there is reunion in fluorescence.When
The concentration of dopamine is 1 mg/ milliliters and during 2 mg/ milliliters, and relatively uniform, but 1mg/ milliliters are distributed phosphor dot quantity more
Fluorescent brightness of the fluorescent brightness less than 2mg/ milliliters.When its concentration is 4mg/ milliliters, the uneven fluorescence bright spot of bulk is formed,
Find after contrast that optimum dopamine concentration is 2mg/ milliliters.
Embodiment 5
1st, the preparation of bionic nano thin film：
A, carrier pretreatment 25mm × (volume ratio is ammonia to 25mm optics microscope slides ammonia washing liquid: hydrogen peroxide: water=1: 1
: ultrapure water is used after 5) soaking 1h, then (volume ratio is hydrochloric acid: hydrogen peroxide: water=1: 1: 5) to be immersed in hydrochloric acid solution
Ultrapure water, N are used after middle 1h2Dry up；
B, dopamine-hydrochloric acid solution is prepared：2mg dopamine-hydrochloric acid is accurately weighed, is dissolved completely in 1 milliliter of 0.1MpH8.5's
Dopamine-the hydrochloric acid solution of 2mg/ milliliters is prepared in Tirs-HCl buffer；
C, will be added in dopamine-hydrochloric acid solution through the microscope slide of pretreatment, temperature（70℃）, thermostatic control oscillator vibration
After 70rpm shakes 1 h, rinsed to face without unnecessary poly-dopamine, N with high purity water2Dry up.
2nd, the fixing meanss of fluorescent probe：Probe is Fluorescein isothiocyanate（FITC）5,20 terminal modified amino length
DNA, be linked to poly-dopamine modification slide on concrete grammar it is as follows,
The preparation of 9.0 carbonate buffer solution of A, 0.5M, pH：0.6g natrium carbonicum calcinatums and 3.7g sodium bicarbonate are dissolved in into 80 respectively
After in ml deionized water, constant volume to 100 milliliters；
B, fluorescent probe solution are prepared：Take during 10 microgram fluorescent probes are dissolved in 1 milliliter of carbonate buffer solution be made into 10 micrograms/
The solution of milliliter；
The link of C, fluorescent probe：The slide that poly-dopamine is modified is dipped in fluorescent probe solution, is shaken in the shaking table of 70rpm
After shaking 4h, deionized water is repeatedly rinsed, and washes unconjugated fluorescent probe off.
Embodiment 6-10：Other steps of embodiment 6-10 change with embodiment 5, only in the preparation of poly-dopamine nano thin-film
The control of polymerization time, fluorescence imaging such as Fig. 5 are carried out to the fluorescence bright spot quantity of above-mentioned four kinds of modes using Image J softwares
Statistical analysiss, as a result such as table 3., there is not fluorescence in the slide of 1h process.There is the fluorescence of several bright spots in the slide of 4h process.When
To 8h, fluorescence bright spot is more than 4h for time lengthening, but still few many than 12h.When the time be 48 h, its fluorescence intensity and
Bright spot quantity is 24h with the suitable therefore optimal polymerization time of 24 h.
Embodiment 11
1st, the preparation of bionic nano thin film：
A, carrier pretreatment 25mm × (volume ratio is ammonia to 25mm optics microscope slides ammonia washing liquid: hydrogen peroxide: water=1: 1
: ultrapure water is used after 5) soaking 1h, then (volume ratio is hydrochloric acid: hydrogen peroxide: water=1: 1: 5) to be immersed in hydrochloric acid solution
Ultrapure water, N are used after middle 1h2Dry up；
B, dopamine-hydrochloric acid solution is prepared：2mg dopamine-hydrochloric acid is accurately weighed, is dissolved completely in 1 milliliter of 0.1MpH8.5's
2% dopamine-hydrochloric acid solution is prepared in Tirs-HCl buffer；
C, will be added in dopamine-hydrochloric acid solution through the microscope slide of pretreatment, temperature（50℃）, thermostatic control oscillator vibration
After 70rpm shaking 24h, rinsed to face without unnecessary poly-dopamine, N with high purity water2Dry up.
2nd, the fixing meanss of fluorescent probe：Probe is Fluorescein isothiocyanate（FITC）5,20 terminal modified amino length
Single stranded DNA, be linked to poly-dopamine modification slide on concrete grammar it is as follows：
The preparation of A, 0.5 M, pH9.0 carbonate buffer solution：0.6g natrium carbonicum calcinatums and 3.7g sodium bicarbonate are dissolved in into 80 respectively
After in ml deionized water, constant volume to 100 milliliters；
B, fluorescent probe solution are prepared：Take during 10 microgram fluorescent probes are dissolved in 1 milliliter of carbonate buffer solution and be made into 10 mcg/mls
Solution；
The link of C, fluorescent probe：The slide that poly-dopamine is modified is dipped in fluorescent probe solution, is shaken in the shaking table of 70rpm
After shaking 1h, deionized water is repeatedly rinsed, and washes unconjugated fluorescent probe off.
Embodiment 12-14：Step changes the probe set time in step C with embodiment 11, only, fluorescence imaging such as Fig. 6,
Statistical analysiss are carried out to the fluorescence bright spot quantity of above-mentioned four kinds of modes using Image J softwares, as a result such as table 4.
Embodiment 11-14 is the optimization of probe rigid condition, and mainly the probe set time is optimized, and fluorescence imaging is seen
Examine and understand（Fig. 6）, the 1h set times, only there are several fluorescence bright spots, show that fixed effect is poor.After 2h, bright spot quantity is
Increase, but effect is still not satisfactory.Extend the set time to after 4h, in four different time sections, it is glimmering that the time point is fixed
Light point quantity is most, and bright spot is evenly distributed, and brightness is most strong.After the time is to 8 h, bright spot quantity is reduced, and low intensity, portion
Divide cluster of grains fusion big, the poly-dopamine thin film for being primarily due to be formed occurs rupture during shaking, affects probe
Fixed effect, therefore Best Times are 4h.
Embodiment 15,（The present embodiment is most preferred embodiment）.
1st, the preparation of bionic nano thin film：
The pretreatment of A, carrier：25mm × 25mm optics microscope slide ammonia washing liquids (volume ratio is ammonia: hydrogen peroxide: water=1:
1: use ultrapure water after 5) soaking 1h, then (volume ratio is hydrochloric acid: hydrogen peroxide: water=1: 1: 5) to be immersed in hydrochloric acid solution
Ultrapure water, N are used after middle 1h2Dry up；
B, dopamine-hydrochloric acid solution is prepared：2mg dopamine-hydrochloric acid is weighed accurately, the Tirs-HCl for being dissolved completely in pH8.5 delays
Rush dopamine-hydrochloric acid solution that 2mg/ milliliters are prepared in liquid；
C, will be added in dopamine-hydrochloric acid solution through the microscope slide of pretreatment, temperature（60℃）, thermostatic control oscillator vibration 70
After rpm shaking 24h, rinsed to face without unnecessary poly-dopamine, N with high purity water2Dry up.
2nd, the fixing meanss of fluorescent probe：Probe is Fluorescein isothiocyanate（FITC）5,20 terminal modified amino length
DNA, be linked to poly-dopamine modification slide on concrete grammar it is as follows：
The preparation of A, 0.5 M pH, 9.0 carbonate buffer solutions：0.6g natrium carbonicum calcinatums and 3.7g sodium bicarbonate are dissolved in into 80 respectively
After in ml deionized water, constant volume to 100 milliliters.
B, fluorescent probe solution are prepared：Take during 10 microgram fluorescent probes are dissolved in 1 milliliter of carbonate buffer solution and be made into 10
The solution of mcg/ml.
The link of C, fluorescent probe：The slide that poly-dopamine is modified is dipped in fluorescent probe solution, shaking in 70rpm
After bed shaking 1-8h, deionized water is repeatedly rinsed, and washes unconjugated fluorescent probe off.
Claims (8)
1. a kind of preparation method of bionic nano thin film, it is characterised in that comprise the steps of：
The pretreatment of A, carrier：Ultrapure water is used after optics microscope slide is soaked with ammonia washing liquid, then is immersed in hydrochloric acid solution
In, then with ultrapure water, N2Dry up；
B, dopamine-hydrochloric acid solution is prepared：Dopamine-hydrochloric acid is weighed, is configured in being dissolved completely in Tirs-HCl buffer
Dopamine-the hydrochloric acid solution of 0.5-4 mg/ milliliters；
C, will be added in dopamine-hydrochloric acid solution through the microscope slide of pretreatment, shake in 50-70 DEG C of thermostatic control oscillator vibration
After shaking 1-48h, rinsed with high purity water, N2Dry up.
2. the preparation method of bionic nano thin film according to claim 1, it is characterised in that：In step A, ammonia is washed
Liquid is made up of the ammonia and hydrogen peroxide and water that volume ratio is 1: 1: 5.
3. the preparation method of bionic nano thin film according to claim 1 and 2, it is characterised in that：Hydrochloric acid in step A
Solution is made up of the hydrochloric acid, hydrogen peroxide and water that volume ratio is 1: 1: 5.
4. the preparation method of bionic nano thin film according to claim 3, it is characterised in that：Dopamine in step B-
Hydrochloric acid solution process for preparation is configured in being dissolved completely in the Tirs-HCl buffer that pH value is 8.5 to weigh dopamine-hydrochloric acid
Dopamine-the hydrochloric acid solution of 2mg/ milliliters.
5. the preparation method of bionic nano thin film according to claim 4, it is characterised in that：When shaking in step C
Between be 24h.
6. the preparation method of bionic nano thin film according to claim 5, it is characterised in that：Thermostatted water in step C
The temperature of bath is 60 DEG C, and the speed of thermostatic control oscillator vibration is 70 rpm.
7. the method for fixing probe with thin film obtained in claim 1, it is characterised in that：Probe is Fluorescein isothiocyanate 5,End
The single stranded DNA of 20 amino length of modification, is linked on the microscope slide of poly-dopamine modification, and concrete grammar is as follows：
The preparation of A, carbonate buffer solution：0.6g natrium carbonicum calcinatums and 3.7g sodium bicarbonate are dissolved in into 80 milliliters of deionizations respectively
After in water, constant volume to 100 milliliters；
B, fluorescent probe solution are prepared：Take during 10 jig probes are dissolved in 1 milliliter of carbonate buffer solution and be made into 10 mcg/mls
Solution；
The link of C, probe：The slide that poly-dopamine is modified is dipped in fluorescent probe solution, is shaken in the shaking table of 70 rpm
After 1-8h, deionized water is rinsed, and washes unconjugated fluorescent probe off.
8. the method that thin film according to claim 7 fixes probe, it is characterised in that：When in step C, shaking table shakes
Between be 4h.
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| CN108051588A (en) * | 2017-12-01 | 2018-05-18 | 美康生物科技股份有限公司 | For the antibody fixing means on the micro-fluidic chip of whole blood sample separation detection |
| CN108126766A (en) * | 2017-12-18 | 2018-06-08 | 深圳市梅丽纳米孔科技有限公司 | A kind of micro-fluidic chip surface hydrophilic method of modifying |
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| CN107383871B (en) * | 2017-07-21 | 2020-03-20 | 安徽师范大学 | Preparation and application of fluorescein isothiocyanate modified poly-dopamine composite material loaded with glucose oxidase |
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| CN111175257A (en) * | 2018-11-13 | 2020-05-19 | 香港城市大学深圳研究院 | Reversible quinone-based modified biochip and preparation method and application thereof |
| CN118032732A (en) * | 2024-04-11 | 2024-05-14 | 大连理工大学 | Method for detecting MPs by covalent chemical labeling method |
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