CN103147133B - Three-dimensional carrier of microarray biochip and preparation method thereof - Google Patents
Three-dimensional carrier of microarray biochip and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a three-dimensional carrier of a microarray biochip and a preparation method thereof. The three-dimensional carrier of a microarray biochip is prepared by growing a zinc oxide nanorod on the surface of a substrate material and growing a polymer brush on the surface of the zinc oxide nanorod. The preparation method comprises the following steps of: activating the surface of the substrate material, and growing a zinc oxide nanorod on the surface of the substrate material; performing surface modification on the grown zinc oxide nanorod; connecting the modified surface of the zinc oxide nanorod with an initiator; and finally, growing a polymer brush to obtain a three-dimensional carrier of a microarray biochip. The preparation method is simple, does not need special equipment, has low production cost and good repeatability, and can be used for realizing large-scale production; the prepared three-dimensional carrier has low price and high fixing density for probe molecule, can be used for effectively inhibiting non-specific protein adsorption and improving the sensitivity and the specificity, and can be widely applied to the fields of biology, basic medicine, disease diagnosis, drug screening, food safety, environmental monitoring and the like.
Description
Technical field
The invention belongs to detection field, relate to the three-dimensional carrier of micro-array biochip, also relate to the preparation method of this three-dimensional carrier.
Background technology
Micro-array biochip be by different bioprobe molecules as DNA, antigen, antibody, aptamers etc. in order, be fixed on certain carrier high-density, addressable, coordinate with fluorescent reading system, realize a kind of technology of extensive, high throughput testing biomolecules and research bio-molecular interaction, at the early screening of disease, Diagnosis and Treat and drug development, basic life science research important in inhibiting, obtain academia in recent years and industry member studies interest widely.At present, the slide glass that the solid support material that micro-array biochip adopts is mainly surface-functionalized, the subject matter that this kind of solid support material exists is limited to the limited surface-area in slide glass one dimension surface, its probe molecule constant density is low, cause the sensitivity of micro-array biochip not high, detectability does not reach application request sometimes.For this problem, some macro-organism medical company have developed several high performance carrier, as the ONCYTE chip of GRACE BIO-LABS company, the PATH chip of GENTEL BIOSCIENCES company, the FAST chip of WHATMAN company, the SUPERPROTEIN chip of ARRAYIT company, the HYDROGEL chip of PERKIN-ELMER company, the MAXISORP chip of NALGE NUNC company, the hydrogel carrier etc. of XanTec analytics, the common ground of these solid support materials is by adopting the three-dimensional carrier with surface micro-structure, to improve the constant density of probe molecule, suppress the non-specific adsorption of biomolecules on surface simultaneously, to obtaining high detection sensitivity.But these commercial chips all exist different defects, such as, need complicated surface active, price is high, poor reproducibility, background signal are high, is difficult to the needs meeting scientific research and practical application.
Summary of the invention
In view of this, an object of the present invention is the three-dimensional carrier providing a kind of micro-array biochip, there is the effect that price is low, high, the effective suppression nonspecific proteins of the constant density of probe molecule adsorbs, can meet in scientific research and practical application sensitivity and specific requirement; Two of object of the present invention is the preparation method providing micro-array biochip three-dimensional carrier, and preparation method is simple.
For achieving the above object, the invention provides following technical scheme:
1. micro-array biochip three-dimensional carrier, described micro-array biochip three-dimensional carrier is at substrate material surface growing zinc oxide nanorod, then obtains at zinc oxide nano rod surface growth polymer brush.
Base material of the present invention can be sheet glass, plastic sheet, nylon membrane, silicon chip etc., and preferably, described base material is slide glass.
As long as have polymer brush in zinc oxide nano rod surface growth in the present invention, described polymer brush is preferably poly-(oligomeric ethylene glycol methyl acrylate-methyl glycidyl acrylate).
In the present invention, polymer brush can use the method such as self-organization, spin coating to grow, and preferably, described polymer brush adopts surperficial Atom Transfer Radical Polymerization technology growth.
2. described in, the preparation method of micro-array biochip three-dimensional carrier, comprises the steps:
A. the surface of base material is activated, then at substrate material surface growing zinc oxide nanorod;
B. the zinc oxide nano rod of growth is carried out finishing;
C. the zinc oxide nano rod surface after modification is connected into initiator;
D. being connected into the zinc oxide nano rod surface growth polymer brush of initiator, the three-dimensional carrier of micro-array biochip is obtained.
Preferably, in described step a, described activation is that 1-10mM potassium permanganate soaks 15-30 minute by base material concentration.
Preferably, in described step a, described growing zinc oxide nanorod is the mixing solutions put into by the base material of activation containing thanomin, ammoniacal liquor and zinc nitrate, 30-60 minute is grown in 65-95 DEG C of water-bath, in described mixing solutions, the final concentration of zinc nitrate is 10-100mM, the volume fraction of thanomin is 1-10%, and the volume fraction of ammoniacal liquor is 1-10%.
Preferably, described step b be by step a process after base material be immersed in containing 3-glycidoxypropyl trimethoxysilane ethanolic soln in soak 1-3 hour, then with after alcohol flushing, vacuum annealing 1-3 hour at 110 DEG C of temperature; Described is 1%-5% containing the volumetric concentration of 3-glycidoxypropyl trimethoxysilane in the ethanolic soln of 3-glycidoxypropyl trimethoxysilane.
Preferably, described step c be by step b process after base material put into containing the tetrahydrofuran solution of triethylamine and α-bromine isobutyl acylbromide or dichloromethane solution 2 hours, by tetrahydrofuran (THF) or dichloromethane rinse after taking-up; In described tetrahydrofuran solution, the volume fraction of triethylamine is the volume fraction of 0.2-0.4%, α-bromine isobutyl acylbromide is 0.4-0.6%.
Preferred, described steps d is that the zinc oxide nano rod surface being connected into initiator is adopted surperficial Atom Transfer Radical Polymerization technology growth polymer brush.Concrete steps are that first preparation is containing 10-30%(v/v) oligomeric ethylene glycol methacrylic ester (molecular-weight average 360) and the methanol/water solution of glycidyl methacrylate 0.4-1%(v/v), wherein the methyl alcohol of methanol/water solution and the volume ratio of water are 1:1, then in solution, nitrogen 15-30 minute is blasted, add 2 again, 2'-dipyridyl and cuprous bromide, to 2, the final concentration of 2'-dipyridyl is 3mg/mL, the final concentration of cuprous bromide is 1.7mg/mL's, after ultrasonic dissolution, slide glass after step c process is immersed rapidly, sealing solution and insert lucifuge inert atmosphere case preserve 6-12 hour.Wherein pass into nitrogen can replace with and add the aqueous ascorbic acid that the concentration being equivalent to liquor capacity 1/50 is 120 mg/mL.
Beneficial effect of the present invention is: the invention discloses micro-array biochip three-dimensional carrier and corresponding preparation method, adopt wet chemical method and surperficial Atom Transfer Radical Polymerization technology in preparation, form the composite structure of zinc oxide nano rod and polymer brush at substrate material surface; The nano structure of zinc oxide formed has huge specific surface area, effectively can improve the surface density of fixing biological probe molecule, and simultaneous oxidation zinc nanometer rod has the specific physical properties amplifying fluorescent signal, greatly improves the signal to noise ratio of fluorometric analysis; And polymer brush is as decorative layer, biomolecules in testing process can be effectively suppressed in the non-specific adsorption on surface, to ensure that highly selective detects in the solution on the one hand; On the other hand owing to containing micro-epoxide group (GMA monomer carries) in polymer brush, in dry conditions can with the amino generation covalent cross-linking of bioprobe molecule, thus realize high-density, high reactivity probe is fixed, therefore detection sensitivity is high, and lowest detectable limit reaches 100pg/mL; The three-dimensional carrier of the micro-array biochip of acquisition can be applied in fundamental research and practical application, play a significant role.
Accompanying drawing explanation
In order to make object of the present invention, technical scheme and beneficial effect clearly, the invention provides following accompanying drawing and being described:
Fig. 1 is micro-array biochip three-dimensional carrier preparation method and polymer brush schematic arrangement.
Fig. 2 is the electron scanning micrograph of zinc oxide nano rod.
Fig. 3 is the electron scanning micrograph of micro-array biochip three-dimensional carrier.
Fig. 4 is the micro-array biochip three-dimensional carrier transmission electron microscope photo of zinc oxide nano rod-polymer brush composite structure.
Fig. 5 is the infared spectrum (a: be zinc oxide nano rod infared spectrum figure of zinc oxide nano rod and zinc oxide nano rod-polymer brush composite structure; B: the infared spectrum of zinc oxide nano rod-polymer brush composite structure).
Fig. 6 is that (a is epoxy activation slide glass for the fluorescence photo of the micro-array chip of 0.8mg/mL fluorescin (CY3 mark anti goat igg) on different carriers, fluorescence intensity and micro-spot diameter; B is that polymer brush modifies slide glass; C is that epoxy active oxidation zinc bar modifies slide glass; D is oxidation zinc bar-polymer brush modification slide glass; Scale in fluorescence photo is 0.5 mm; ).
Fig. 7 is the fluorescence photo that micro-array biochip three-dimensional carrier soaks before and after 30 minutes in 5 mcg/ml fluorescins (CY3-anti goat igg) solution.
Fig. 8 is that micro-array biochip three-dimensional carrier adopts 10% serum sample of micro-array chip to tumor markers CEA to detect the fluorescence photo and typical curve that obtain.
Embodiment
Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are described in detail.
Micro-array biochip three-dimensional carrier, its structure and preparation technology are as shown in Figure 1.
As shown in Figure 1, micro-array biochip three-dimensional carrier by the zinc oxide nano rod of substrate material surface growing high density, random orientation, then obtains at zinc oxide nano rod surface growth polymer brush.Growing polymer brush can pass through surperficial Atom Transfer Radical Polymerization (Surface-initiated Atom Transfer Radical Polymerization, SI-ATRP) technology.
embodiment 1
The preparation method of micro-array biochip three-dimensional carrier, concrete steps are as follows:
A. clean slide is immersed in 5 mM and newly prepares potassium permanganate solution 20 minutes, taking-up deionized water rinsing; Then slide glass is put into containing 5%(v/v) thanomin and 5%(v/v) 50 mM zinc nitrate solutions of ammoniacal liquor, 50 minutes are grown under 85 DEG C of water-baths, slide surface is made to form zinc oxide nano rod, dry after taking-up deionized water rinsing, then observe with scanning electron, result as shown in Figure 2;
B. the slide glass after step a process is immersed in the ethanolic soln containing 3% (v/v) 3-glycidoxypropyl trimethoxysilane (APTES) and soaks 2 hours, 3-glycidoxypropyl trimethoxysilane and zinc oxide nano rod is made to form silicon-oxygen covalent linkage, with modification zinc oxide nanorod surfaces, after alcohol flushing, vacuum annealing 2 hours at 110 DEG C of temperature;
C. the slide glass after step b process is put into containing 0.35%(v/v) triethylamine (triethylamine, TEA) α and 0.6%(v/v)-bromine isobutyl acylbromide (2-bromoisobutyryl bromide, BIB) soak 2 hours in dichloromethane solution, dichloromethane rinse three times are used, to be connected into initiator on zinc oxide nano rod surface after taking out;
D. growing polymer brush on the slide glass being adopted by the slide glass through step c process surperficial Atom Transfer Radical Polymerization technology to have zinc oxide in this growth, be specially: first prepare 50 milliliters containing 10%(v/v) oligomeric ethylene glycol methacrylic ester (oligo(ethylene glycol) methacrylate, OEGMA, molecular-weight average 360) and 0.4%(v/v) glycidyl methacrylate (glycidyl methacrylate, GMA) methanol/water (volume ratio is 1:1) solution, nitrogen 15-30 minute is blasted in solution, add 150 milligram 2 again, 2'-dipyridyl (2, 2'-Dipyridyl) He 70 milligrams of cuprous bromides, after ultrasonic dissolution, the slide glass of step c process is immersed rapidly, sealing solution the inert atmosphere case of inserting lucifuge preserves 12 hours, obtain micro-array biochip three-dimensional carrier.Gained micro-array biochip three-dimensional carrier is observed under scanning electronic microscope and transmission electron microscope, result as shown in Figure 3 and Figure 4, from Fig. 3 and Fig. 4, at gained micro-array biochip three-dimensional carrier, there is irregular zinc oxide nano rod in slide surface growth, and define polymer brush on zinc oxide nano rod surface.
The three-dimensional carrier of the slide glass in conjunction with zinc oxide nano rod prepared by step c and steps d gained micro-array biochip carries out infared spectrum analysis, and result as shown in Figure 5.Have polymer brush by infared spectrum is known in the surface growth of oxidation nanometer rod, in the present embodiment, polymer brush is poly-(oligomeric ethylene glycol methyl acrylate-methyl glycidyl acrylate).
embodiment 2
The preparation method of micro-array biochip three-dimensional carrier, concrete steps are as follows:
A. clean slide is immersed in 1 mM and newly prepares potassium permanganate solution 30 minutes, taking-up deionized water rinsing; Then slide glass is put into containing 1%(v/v) thanomin and 1%(v/v) 10 mM zinc nitrate solutions of strong aqua, in 65 DEG C of water-baths, grow 60 minutes, make slide surface form zinc oxide nano rod, dry after taking-up deionized water rinsing;
B. the slide glass after step a process is immersed in containing 1%(v/v) 3-glycidoxypropyl trimethoxysilane (APTES) ethanolic soln in soak 3 hours, 3-glycidoxypropyl trimethoxysilane and zinc oxide nano rod surface is made to form silicon-oxygen covalent linkage, with modification zinc oxide nanorod surfaces, with after alcohol flushing at 110 DEG C of temperature, vacuum annealing 3 hours;
C. the slide glass after step b process is put into containing 0.2%(v/v) triethylamine (triethylamine, TEA) and 0.4%(v/v) α-bromine isobutyl acylbromide (2-bromoisobutyryl bromide, BIB) soak 2 hours in tetrahydrofuran solution, three times are rinsed with tetrahydrofuran (THF), to be connected into initiator on zinc oxide nano rod surface after taking-up;
D. growing polymer brush on the slide glass being adopted by the slide glass through step c process surperficial Atom Transfer Radical Polymerization technology to have zinc oxide in this growth, be specially: first prepare 50 milliliters containing 30%(v/v) oligomeric ethylene glycol methacrylic ester (oligo (ethylene glycol) methacrylate, OEGMA, molecular-weight average 360) and 1%(v/v) glycidyl methacrylate (glycidyl methacrylate, GMA) methanol/water (volume ratio is 1:1) solution, 230 milligram 2 is added again in solution, 2'-dipyridyl (2, 2 '-bipyridyl) and 170 milligrams of cupric bromides, then after the slide glass of step c process being immersed, add rapidly the aqueous ascorbic acid that 1mL concentration is 120mg/mL, wherein 2, 2'-dipyridyl and cupric bromide form complex compound Cu(II in the solution)-2Bpy, this complex compound is reduced to cuprous complex Cu(I by xitix)-2Bpy, then at surface catalysis Atom Transfer Radical Polymerization.Seal and the solution that vibrates, the inert atmosphere case of then inserting lucifuge preserves 12 hours, obtains the three-dimensional carrier of micro-array biochip.
embodiment 3
The preparation method of the three-dimensional carrier of micro-array biochip, concrete steps are as follows:
A. clean slide is immersed in 10 mM and newly prepares potassium permanganate solution 15 minutes, taking-up deionized water rinsing; Then slide glass is put into containing 0.25%(v/v) thanomin and 5%(v/v) 100 mM zinc nitrate solutions of strong aqua, 60 minutes are grown in 95 DEG C of water-baths, slide surface is made to form zinc oxide nano rod, dry after taking-up deionized water rinsing, then observe with scanning electron, result as shown in Figure 2;
B. the slide glass after step a process is immersed in containing 5%(v/v) soak 1 hour in the ethanolic soln of 3-glycidoxypropyl trimethoxysilane (APTES), 3-glycidoxypropyl trimethoxysilane and zinc oxide nano rod surface is made to form silicon-oxygen covalent linkage, with modification zinc oxide nanorod surfaces, with vacuum annealing 1 hour at 110 DEG C of temperature after alcohol flushing;
C. the slide glass after step b process is put into containing 0.4%(v/v) triethylamine (triethylamine, TEA) and 0.4%(v/v) α-bromine isobutyl acylbromide (2-bromoisobutyryl bromide, BIB) soak 2 hours in dichloromethane solution, dichloromethane rinse three times are used, to be connected into initiator on zinc oxide nano rod surface after taking out;
D. growing polymer brush on the slide glass being adopted by the slide glass after step c process surperficial Atom Transfer Radical Polymerization technology to have zinc oxide in this growth, be specially: first prepare 50 milliliters containing 20%(v/v) oligomeric ethylene glycol methacrylic ester (oligo(ethylene glycol) methacrylate, OEGMA, molecular-weight average 360) and glycidyl methacrylate (the glycidyl methacrylate of 0.8% (v/v), GMA) methanol/water (volume ratio is 1:1) solution, nitrogen 15-30 minute is blasted in solution, add 150 milligram 2 again, 2'-dipyridyl (2, 2 '-Dipyridyl) and 70 milligrams of cuprous bromides, after ultrasonic dissolution, the slide glass of step c gained in conjunction with zinc oxide nano rod is immersed rapidly, sealing solution the inert atmosphere case of inserting lucifuge preserves 10 hours, obtain the three-dimensional carrier of micro-array biochip.
embodiment 4
Biochip preparation and detecting step:
The anti goat igg (anti-Goat IgG) that probe molecule CY3 marks is dissolved in 0.01M PBS solution, make the solution that concentration is 800 μ g/mL, adopt the three-dimensional carrier surface printing microarray that micro-array chip point sample instrument (contact point sample or contactless spot sample mode) is prepared in embodiment 1, after point sample, three-dimensional carrier is left standstill 8-12 hour at room temperature 20-25 DEG C, carry out fluorescent scanning with after the cleaning of 0.01M PBS solution, drying again, scanning result as shown in fig 6d.Then adopt the microarray that same solution prints at other carrier surfaces, then carry out fluorescent scanning, wherein Fig. 6 a is at epoxy activation slide surface printing microarray; Fig. 6 b modifies slide surface printing microarray at poly-(oligomeric ethylene glycol methyl acrylate-methyl glycidyl acrylate) polymer brush; Fig. 6 c is the microarray modifying slide surface printing at epoxy active oxidation zinc bar.Compared with microarray prepared by the microarray prepared by the three-dimensional carrier that the present invention obtains and other carriers, the fluorescence intensity that three-dimensional carrier prepared by the present invention obtains obviously strengthens, and each micro-some even intensity, shape specification.Therefore, use three-dimensional carrier of the present invention that protein probe molecular density can be made to be fixed on three-dimensional carrier surface, be conducive to the detection of micro-array chip.
The three-dimensional carrier of micro-array biochip embodiment 1 prepared is submergence 30 minutes in the anti goat igg solution that marks of 5 μ g/mLCY3 in concentration, cleans, carries out fluorescent scanning after drying, obtain fluorescence photo shown in Fig. 7 b after taking out by 0.01M PBS solution.(Fig. 7 a), the fluorescence intensity of this three-dimensional carrier, without obvious rising, demonstrates three-dimensional carrier prepared by the present invention and under solution condition, is highly resistant to nonspecific proteins absorption, for highly selective immunodetection provides guarantee to compare the fluorescence photo before immersion.
Probe molecule anti-carcinoembryonic antigen monoclonal antibody (monoclonal anti-CEA) is dissolved in 0.01M PBS solution, make the solution that concentration is 800 μ g/mL, the three-dimensional carrier surface printing microarray of the micro-array biochip adopting micro-array chip point sample instrument to prepare in embodiment 1, after point sample by three-dimensional carrier at room temperature leave standstill 8-12 hour, then with 0.01M PBS solution cleaning, drying.Then adopt sandwich assay to tumor markers carcinomebryonic antigen (carcino-embryonic antigen, CEA) detect, namely first chip is reacted 1 hour with the 10% human serum solution soaking containing different concns CEA respectively, again with polyclonal antibody (the polyclonal anti-CEA of anti-carcinoembryonic antigen, produced in Mouse) react 30 minutes, last and CY3 marks against murine IgG and reacts 15 minutes, dry after cleaning, then adopt Fluorescence Scanner to read chip signal, obtain fluorescence photo shown in Fig. 8 a.The intensity of fluorescent signal is used for drawing standard curve, as shown in Figure 8 b.Obtained in human serum, can reaching 100pg/mL based on the micro-array chip of this three-dimensional carrier to the detectability of CEA by typical curve, therefore detection sensitivity is high, has excellent performance.
What finally illustrate is, above preferred embodiment is only in order to illustrate technical scheme of the present invention and unrestricted, although by above preferred embodiment to invention has been detailed description, but those skilled in the art are to be understood that, various change can be made to it in the form and details, and not depart from claims of the present invention limited range.
Claims (2)
1. the three-dimensional carrier of micro-array biochip, it is characterized in that: the three-dimensional carrier of described micro-array biochip is at substrate material surface growing zinc oxide nanorod, obtain at zinc oxide nano rod surface growth polymer brush, described polymer brush is poly-(oligomeric ethylene glycol methyl acrylate-methyl glycidyl acrylate) again;
Comprise the steps:
A. the surface of base material is activated, then at substrate material surface growing zinc oxide nanorod; Described activation is that 1-10mM potassium permanganate soaks 15-30 minute by base material concentration; Described growing zinc oxide nanorod is the mixing solutions put into by the base material of activation containing thanomin, ammoniacal liquor and zinc nitrate, 30-60 minute is grown in 65-95 DEG C of water-bath, in described mixing solutions, the final concentration of zinc nitrate is 10-100mM, the volume fraction of thanomin is 1-10%, and the volume fraction of ammoniacal liquor is 1-10%
B. the base material after step a process is immersed in the ethanolic soln containing 3-glycidoxypropyl trimethoxysilane and soaks 1-3 hour, then with after alcohol flushing, vacuum annealing 1-3 hour at 110 DEG C of temperature; Described is 1%-5% containing the volumetric concentration of 3-glycidoxypropyl trimethoxysilane in the ethanolic soln of 3-glycidoxypropyl trimethoxysilane;
C. the base material after step b process is put into containing the tetrahydrofuran solution of triethylamine and α-bromine isobutyl acylbromide or dichloromethane solution 2 hours, by tetrahydrofuran (THF) or dichloromethane rinse after taking-up; In described tetrahydrofuran solution, the volume fraction of triethylamine is the volume fraction of 0.2%-0.4%, α-bromine isobutyl acylbromide is 0.4%-0.6%;
D. the zinc oxide nano rod surface being connected into initiator is adopted surperficial Atom Transfer Radical Polymerization technology growth polymer brush, obtain the three-dimensional carrier of micro-array biochip.
2. micro-array biochip three-dimensional carrier according to claim 1, is characterized in that: described base material is sheet glass, plastic sheet, nylon membrane or silicon chip.
3. the preparation method of micro-array biochip three-dimensional carrier described in any one of claim 1 to 2, is characterized in that, comprise the steps:
A. the surface of base material is activated, then at substrate material surface growing zinc oxide nanorod; Described activation is that 1-10mM potassium permanganate soaks 15-30 minute by base material concentration; Described growing zinc oxide nanorod is the mixing solutions put into by the base material of activation containing thanomin, ammoniacal liquor and zinc nitrate, 30-60 minute is grown in 65-95 DEG C of water-bath, in described mixing solutions, the final concentration of zinc nitrate is 10-100mM, the volume fraction of thanomin is 1-10%, and the volume fraction of ammoniacal liquor is 1-10%;
B. the base material after step a process is immersed in the ethanolic soln containing 3-glycidoxypropyl trimethoxysilane and soaks 1-3 hour, then with after alcohol flushing, vacuum annealing 1-3 hour at 110 DEG C of temperature; Described is 1%-5% containing the volumetric concentration of 3-glycidoxypropyl trimethoxysilane in the ethanolic soln of 3-glycidoxypropyl trimethoxysilane;
C. the base material after step b process is put into containing the tetrahydrofuran solution of triethylamine and α-bromine isobutyl acylbromide or dichloromethane solution 2 hours, by tetrahydrofuran (THF) or dichloromethane rinse after taking-up; In described tetrahydrofuran solution, the volume fraction of triethylamine is the volume fraction of 0.2%-0.4%, α-bromine isobutyl acylbromide is 0.4%-0.6%;
D. the zinc oxide nano rod surface being connected into initiator is adopted surperficial Atom Transfer Radical Polymerization technology growth polymer brush, obtain the three-dimensional carrier of micro-array biochip.
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| CN104034884B (en) * | 2014-07-01 | 2016-08-24 | 西南大学 | Preparation method of microarray antibody chip based on dendritic polymer and products thereof |
| CN104761745B (en) * | 2015-03-16 | 2017-11-24 | 北京化工大学 | A kind of three-dimensional biochip substrate preparation method |
| CN104880447B (en) * | 2015-06-09 | 2018-02-13 | 吉林大学 | A kind of ZnO NRs/Au films complex three-dimensional micro-array biochip substrate and preparation method thereof |
| CN105255719B (en) * | 2015-10-27 | 2017-10-03 | 武汉大学 | A kind of three-dimensional porous micro-fluidic chip and preparation method and application |
| CN107486270B (en) * | 2017-08-08 | 2020-10-23 | 上海格荣生物科技有限公司 | Preparation method of microarray chip based on ball-brush double-layer nanostructure substrate |
| CN110702642B (en) * | 2019-10-29 | 2022-03-18 | 西南大学 | Preparation method of micro-well structured SPRi chip, product and application thereof |
| CN113447446B (en) * | 2021-06-28 | 2023-05-05 | 西南大学 | Chip for quantitatively detecting reductive multicomponent biological micromolecules by utilizing oblique incident light reflection difference, application and detection method |
| CN114460047A (en) * | 2021-12-20 | 2022-05-10 | 上海中医药大学 | Artificial modified 3D photo-crosslinking probe and method for fishing Chinese herbal medicine active small-molecule synthetase |
| CN114904595B (en) * | 2022-06-21 | 2023-07-25 | 中国科学院长春应用化学研究所 | Substrate microarray chip based on gold nanorod-brush double-layer nanostructure and preparation method thereof |
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