CN103234940A - Molecular imprinting sensing chip available for underwater detection - Google Patents
Molecular imprinting sensing chip available for underwater detection Download PDFInfo
- Publication number
- CN103234940A CN103234940A CN2013101779297A CN201310177929A CN103234940A CN 103234940 A CN103234940 A CN 103234940A CN 2013101779297 A CN2013101779297 A CN 2013101779297A CN 201310177929 A CN201310177929 A CN 201310177929A CN 103234940 A CN103234940 A CN 103234940A
- Authority
- CN
- China
- Prior art keywords
- chip
- molecular
- film
- water
- layers
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000001514 detection method Methods 0.000 title abstract description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 50
- 239000011521 glass Substances 0.000 claims abstract description 34
- 125000000217 alkyl group Chemical group 0.000 claims abstract description 28
- 239000000758 substrate Substances 0.000 claims abstract description 26
- 239000008346 aqueous phase Substances 0.000 claims abstract description 15
- 229910052751 metal Inorganic materials 0.000 claims abstract description 13
- 239000002184 metal Substances 0.000 claims abstract description 13
- 238000006116 polymerization reaction Methods 0.000 claims description 34
- 239000012528 membrane Substances 0.000 claims description 29
- 239000000243 solution Substances 0.000 claims description 25
- 239000007788 liquid Substances 0.000 claims description 24
- 239000000126 substance Substances 0.000 claims description 24
- 238000006243 chemical reaction Methods 0.000 claims description 22
- 229910052804 chromium Inorganic materials 0.000 claims description 21
- 239000011651 chromium Substances 0.000 claims description 21
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 20
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 claims description 20
- 238000000034 method Methods 0.000 claims description 19
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 18
- 230000000977 initiatory effect Effects 0.000 claims description 15
- 238000004132 cross linking Methods 0.000 claims description 14
- 239000012153 distilled water Substances 0.000 claims description 13
- 239000000463 material Substances 0.000 claims description 13
- 239000000376 reactant Substances 0.000 claims description 13
- 239000003480 eluent Substances 0.000 claims description 11
- 238000001704 evaporation Methods 0.000 claims description 11
- 230000008020 evaporation Effects 0.000 claims description 11
- 239000000178 monomer Substances 0.000 claims description 11
- 229920000642 polymer Polymers 0.000 claims description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 10
- 239000004088 foaming agent Substances 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 10
- 229910052757 nitrogen Inorganic materials 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 10
- 238000012360 testing method Methods 0.000 claims description 9
- 238000010276 construction Methods 0.000 claims description 8
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 8
- 239000010931 gold Substances 0.000 claims description 8
- 229910052737 gold Inorganic materials 0.000 claims description 8
- 238000007738 vacuum evaporation Methods 0.000 claims description 8
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 6
- 239000003795 chemical substances by application Substances 0.000 claims description 6
- 230000008649 adaptation response Effects 0.000 claims description 5
- 238000011010 flushing procedure Methods 0.000 claims description 5
- 238000007747 plating Methods 0.000 claims description 5
- 238000001338 self-assembly Methods 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 4
- 230000015572 biosynthetic process Effects 0.000 claims description 3
- 238000007654 immersion Methods 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 239000011259 mixed solution Substances 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- 229920000344 molecularly imprinted polymer Polymers 0.000 abstract description 24
- 239000001257 hydrogen Substances 0.000 abstract description 6
- 229910052739 hydrogen Inorganic materials 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 5
- 230000006378 damage Effects 0.000 abstract description 3
- 238000011896 sensitive detection Methods 0.000 abstract description 2
- 230000001953 sensory effect Effects 0.000 abstract 1
- 239000010408 film Substances 0.000 description 70
- 238000005286 illumination Methods 0.000 description 16
- RJKFOVLPORLFTN-LEKSSAKUSA-N Progesterone Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 RJKFOVLPORLFTN-LEKSSAKUSA-N 0.000 description 12
- MUMGGOZAMZWBJJ-DYKIIFRCSA-N Testostosterone Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 MUMGGOZAMZWBJJ-DYKIIFRCSA-N 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- WNAHIZMDSQCWRP-UHFFFAOYSA-N dodecane-1-thiol Chemical compound CCCCCCCCCCCCS WNAHIZMDSQCWRP-UHFFFAOYSA-N 0.000 description 9
- STVZJERGLQHEKB-UHFFFAOYSA-N ethylene glycol dimethacrylate Chemical class CC(=C)C(=O)OCCOC(=O)C(C)=C STVZJERGLQHEKB-UHFFFAOYSA-N 0.000 description 8
- 230000006870 function Effects 0.000 description 8
- 230000004044 response Effects 0.000 description 8
- 239000010409 thin film Substances 0.000 description 8
- 229960003387 progesterone Drugs 0.000 description 6
- 239000000186 progesterone Substances 0.000 description 6
- DBCAQXHNJOFNGC-UHFFFAOYSA-N 4-bromo-1,1,1-trifluorobutane Chemical compound FC(F)(F)CCCBr DBCAQXHNJOFNGC-UHFFFAOYSA-N 0.000 description 5
- 239000007864 aqueous solution Substances 0.000 description 5
- 229960003604 testosterone Drugs 0.000 description 5
- VOXZDWNPVJITMN-ZBRFXRBCSA-N 17β-estradiol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 VOXZDWNPVJITMN-ZBRFXRBCSA-N 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 239000012295 chemical reaction liquid Substances 0.000 description 4
- 229960005309 estradiol Drugs 0.000 description 4
- 229930182833 estradiol Natural products 0.000 description 4
- RWCCWEUUXYIKHB-UHFFFAOYSA-N benzophenone Chemical compound C=1C=CC=CC=1C(=O)C1=CC=CC=C1 RWCCWEUUXYIKHB-UHFFFAOYSA-N 0.000 description 3
- 239000012965 benzophenone Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 229910052746 lanthanum Inorganic materials 0.000 description 3
- FZLIPJUXYLNCLC-UHFFFAOYSA-N lanthanum atom Chemical compound [La] FZLIPJUXYLNCLC-UHFFFAOYSA-N 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 125000005395 methacrylic acid group Chemical class 0.000 description 3
- 238000009738 saturating Methods 0.000 description 3
- 239000000427 antigen Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000006073 displacement reaction Methods 0.000 description 2
- 239000003792 electrolyte Substances 0.000 description 2
- 230000009471 action Effects 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000008358 core component Substances 0.000 description 1
- 238000007877 drug screening Methods 0.000 description 1
- 238000000835 electrochemical detection Methods 0.000 description 1
- 230000009881 electrostatic interaction Effects 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000006386 memory function Effects 0.000 description 1
- 238000004375 physisorption Methods 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
Images
Landscapes
- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
本发明涉及一种可用于水中检测的分子印迹传感芯片，具体涉及分子印迹薄膜传感芯片领域。具有多层结构；由玻璃基片、金属膜、单分子烷基链层、多层分子印迹膜组成；该芯片按照玻璃基片、金属膜、单分子烷基链层、多层分子印迹膜由下至上的顺序依次排列。由于采用形成多层分子印迹聚合物膜的方法制备了SPR传感器芯片，可以有效的减弱水对分子印迹膜中氢键的破坏作用，从而使合成的分子印迹膜能够在水中存活，并能实现在水相中对模板分子进行灵敏检测的效果，为实际环境中精确检测样品的含量提供了强有力的检测手段。因此可以有效解决传统传感芯片不能在水相中检测样品这一难题。The invention relates to a molecularly imprinted sensor chip that can be used for detection in water, in particular to the field of molecularly imprinted film sensory chips. It has a multi-layer structure; it is composed of a glass substrate, a metal film, a single-molecular alkyl chain layer, and a multi-layer molecularly imprinted film; the chip is composed of a glass substrate, a metal film, a single-molecular alkyl chain layer, and a multi-layer molecularly imprinted film. Arranged in order from bottom to top. Since the SPR sensor chip is prepared by forming a multi-layer molecularly imprinted polymer film, it can effectively weaken the damage effect of water on the hydrogen bonds in the molecularly imprinted film, so that the synthesized molecularly imprinted film can survive in water and can realize The effect of sensitive detection of template molecules in the aqueous phase provides a powerful detection method for accurately detecting the content of samples in the actual environment. Therefore, it can effectively solve the problem that traditional sensor chips cannot detect samples in the aqueous phase.
Description
Technical field
The present invention relates to a kind of molecular engram sensing chip that can be used for detecting in the water, be specifically related to molecule engram film sensing chip field.
Background technology
Molecular imprinting is to come from biological chemistry, and a cross discipline technology of subjects such as materials chemistry and high polymer chemistry belongs to the host-guest chemistry category in the supramolecular chemistry.The theory of antibody-antigen is that the development of molecular imprinting is laid a good foundation in the biology.Antibody theory according to Pauling is described, and molecular imprinting as being similar to antigen " template ", forms the molecularly imprinted polymer that template molecule there be " memory function " with function monomer and crosslinking chemical reaction with template molecule.The preparation molecularly imprinted polymer generally includes three processes: the functional group of (1) function monomer and template molecule is reversible combination under proper condition, forms host-guest complex; (2) add crosslinking chemical and initiating agent, initiation under proper condition, crosslinking chemical and function monomer reactive polymeric, the polymkeric substance of template molecule that formed embedding; (3) with physics or chemical method, wash-out template molecule from polymkeric substance, skeleton formation space configuration and binding site at polymkeric substance all " are remembered " hole that the template molecule characteristics, obtain molecularly imprinted polymer, described molecularly imprinted polymer has specific recognition to template molecule, be that described molecularly imprinted polymer has narrow spectrum selection combination to template molecule
The molecular engram sensing chip is the core component of molecular engram sensor, is one of important directions of molecular imprinting development in recent years, is having a wide range of applications aspect chemistry and the biological detection.The molecular engram sensing chip is fixed on the substrate surface of chip with molecular engram material with film or form of powder usually, and sample molecule is detected.
In the biology sample detection field, compare with the detection technique of routine, surface plasma resonance sensor has the complicated sample of need not enrichment and purification step, need not mark, easy and simple to handle, high sensitivity, response fast and the outstanding advantages such as biologically active that keep molecule, be subjected to people's extensive concern.
Surface plasma resonance is a kind of physical optics phenomenon, it is very responsive to the electrolyte that is attached to the metal surface, all electrolyte that are attached to the metal surface all can be detected, and therefore can study chemistry and the physisorption character of material by the variation of measuring the metal surface material refractive index.At present, the biosensor technique of surface plasma resonance has been widely used in a plurality of Biochemical Research fields such as immunology, drug screening.
Be used for the molecular engram sensing chip of surface plasma resonance sensor, comprise glass substrate, golden film and molecular engram film, wherein molecular engram film is its topmost structure.
In the process of molecular engram film preparation, traditional molecular engram film all adopts the form of individual layer, and is destroyed under the effect of outer field molecular engram film hydrogen bond in water when test sample in water, therefore can not detect in water.The environment of test sample only is confined in the organic solution.And the sample in the practical application mostly exists under the environment of water, this also is that molecule engram film can not apply to a bottleneck problem in the reality.
Summary of the invention
The objective of the invention is to propose a kind of molecular engram sensing chip that can be used for detecting in the water in order to solve the problem that existing molecular engram sensing chip can not detect in water.
The objective of the invention is to be achieved through the following technical solutions:
A kind of molecular engram sensing chip that can be used for detecting in the water of the present invention has sandwich construction; Formed by glass substrate, metal film, unimolecule alkyl chain layer, multilayer molecular blotting membrane; This chip is arranged in order according to glass substrate, metal film, unimolecule alkyl chain layer, multilayer molecular blotting membrane order from the bottom to top.
The preparation method of this sensing chip, concrete steps are as follows:
The method of step 1, employing vacuum evaporation is at glass substrate plating one deck chromium film; Follow evaporation one deck gold film; With the 3 layers of chip of material called after that obtain;
Step 7, configuration eluent carry out wash-out to the multilayer chiop of step 7 gained, remove the template molecule in the molecular engram film, and obtaining can be at the molecular engram sensing chip of aqueous phase test sample;
More stable when detecting in water for the multilayer molecular trace rete chip that makes gained, can before the step 7 wash-out, handle chip, make it adapt to aqueous environment; Disposal route is with distilled water or PBS solution injection reaction tank, to make the aqueous environment in the abundant adaptive response of the resulting multilayer molecular trace of the step 6 rete chip pond;
The amount of substance concentration of solute alkyl sulfhydryl is 1-100mmol/L in the alkyl sulfhydryl solution described in the step 2, and solvent is a kind of in methyl alcohol, ethanol or the methenyl choloride.
4 layers of chip described in the step 2 need to dry up with alcohol flushing and with nitrogen before use.
The described template molecule of step 3 and step 4 is sample to be detected; The mol ratio of template molecule, function monomer and crosslinking chemical is 1:4:3～15; The volumetric molar concentration of template molecule in pore-foaming agent is 0.01～0.1mol/L, and the concentration of function monomer in pore-foaming agent is 0.04～0.4mol/L, and the concentration of crosslinking chemical in pore-foaming agent is 0.03～1.5mol/L; The concentration of initiating agent in pore-foaming agent is 1～10mg/ml;
The number of plies of the molecule engram film that the described multilayer molecular trace of step 6 rete chip forms is 2～5;
The described initiation conditions of step 5 and step 6 is that heat causes, ultraviolet light causes or the electropolymerization method;
Eluent described in the step 7 is the mixed solution of distilled water and acetic acid, and mixed volume is than 9:1～7:3;
Principle of work of the present invention is:
At first, form host-guest complex by hydrogen bond or electrostatic interaction between template molecule and the function monomer molecule, behind adding crosslinking chemical and the initiating agent and behind the initiated polymerization, grow the molecule engram film that one deck contains template molecule in metallic film surface, carry out wash-out with eluent then, eluent is by aqueous phase solution and acetic acid preparation, be high polarity, acting force between it and the template molecule is stronger than the acting force between function monomer and the template molecule, can from molecular engram film, seize template molecule, thereby template molecule is washed off, stay and the template molecule shape, corresponding and contain the hole in selective binding site on the spaces such as size, this cavity energy specific recognition and seizure sample molecule cause change of refractive between molecular engram film and the metal film.
Divide the period of the day from 11 p.m. to 1 a.m in the aqueous phase test sample, owing in the aqueous phase solution, contain hydrogen bond, destroyed the structure of molecule engram film, so traditional molecular engram sensing chip can not apply to aqueous phase.But sensing chip of the present invention is owing to comprise double-deck molecule engram film; outer field molecular engram film shields; protecting hydrogen bond action in the molecular engram film of internal layer not to be subjected to or to be subjected to the destruction of not half; thereby a kind of molecular engram sensing chip with new construction of the present invention can detect the sample molecule of aqueous phase; make simple; good reproducibility can apply in the middle of the commercial production widely.
Beneficial effect:
1, a kind of molecular engram sensing chip that can be used for detecting in the water of the present invention, owing to adopt the method that forms multilayer molecular imprinted polymer film to prepare spr sensor chip, can effectively weaken water to the destruction of hydrogen bond in the molecular engram film, thereby synthetic molecular engram film can be survived in water, and can be implemented in aqueous phase template molecule is carried out the effect of Sensitive Detection, in the actual environment accurately the content of test sample strong detection means is provided.Therefore can effectively solve traditional sensing chip can not be in this difficult problem of aqueous phase test sample.
2, a kind of molecular engram sensing chip that can be used for detecting in the water of the present invention adopts chromium film and two kinds of metal films of golden film to prepare spr sensor chip.During test sample, the chromium film can avoid golden film destroyed in water to sensing chip, plays the protective money film, supports golden film in water like this, makes the more stable effect in water of golden film, thereby makes sensing chip better test sample in water.
3, a kind of molecular engram sensing chip that can be used for detecting in the water of the present invention, make this chip produce response to the sample molecule of aqueous phase, specific recognition is strong, applied widely, can be used for SPR detects, QCM detects and Electrochemical Detection, the thickness of the molecular engram film of chip surface can accurately be controlled by reaction time length, the film of preparation nanoscale and even micron order dimensional thickness, adopt in-situ polymerization technology, avoided pulverizing, fix and wait fixing operation, and this film recycling rate of waterused height, good reproducibility, production cost is low, is conducive to suitability for industrialized production.
Description of drawings
Fig. 1 is the structural representation of sensing chip of the present invention;
Fig. 2 is the SPR curve before and after the prepared sensing chip wash-out among the embodiment 1;
Fig. 3 is for adopting the SPR curve before and after the prepared sensing chip absorption testosterone among the embodiment 1.
Fig. 4 is the SPR curve before and after the prepared sensing chip wash-out among the embodiment 2;
Fig. 5 is for adopting the SPR curve before and after the prepared sensing chip absorption progesterone among the embodiment 2.
Wherein, 1-glass substrate, 2-chromium rete, 3-golden membranous layer, 4-unimolecule alkyl chain layer, 5-first molecular imprinted polymer membrane, 6-second molecular engram film.
Embodiment
Below in conjunction with drawings and Examples content of the present invention is described further.
The new construction of a kind of molecular engram sensing chip of the present invention as shown in Figure 1, comprises glass substrate 1, chromium rete 2, golden membranous layer 3, unimolecule alkyl chain layer 4, first molecular imprinted polymer membrane 5, second molecular imprinted polymer membrane 6; This chip is followed successively by glass substrate 1, chromium rete 2, golden membranous layer 3, unimolecule alkyl chain layer 4, first molecular imprinted polymer membrane 5, second molecular imprinted polymer membrane 6 from lower to upper;
Be template molecule with the testosterone, prepare a kind of molecular engram sensing chip with new construction with light-initiated method, its concrete steps are as follows:
1) adopt the method for vacuum evaporation at glass substrate 1 plating one deck chromium film and one deck gold film; The method of concrete evaporation is: with the chromium that the glass substrate 1 that cleans carries out vacuum evaporation 99.99%, vacuum tightness is 〉=1 * 10
-5Pa, evaporation speed is
The thickness of chromium film is 2nm; The then gold of evaporation 99.99% on chromium film 2, the thickness of gold-plated film be 41nm; The chip that obtain this moment is 3 layers of chip;
Wherein, described glass substrate 1 is the lanthanum glass of 2.5cm * 30cm;
3 layers of chip that 2) will newly plate immerse the ethanolic solution of the lauryl mercaptan of 1mM immediately, soak 24h, soak the back self assembly one deck unimolecule alkyl chain layer 4 on golden membranous layer 3 that finishes, and it is used alcohol flushing, and it is standby that nitrogen dries up the back, obtains 4 layers of chip;
Wherein, described lauryl mercaptan solution is the ethanolic solution of 1mmol/L lauryl mercaptan, and chemical formula is SH (CH
2)
11CH
3
3) 0.028g testosterone and 34 μ L methacrylic acids are dissolved in the 4mL acetonitrile in, ultrasonic mixing 5min leaves standstill 3h; Add again 56.5 μ L ethylene glycol dimethacrylates and with 0.016g benzophenone, ultrasonic mixing 5min, logical nitrogen 10min then, preparation prepolymerization reaction liquid A;
4) with step 3 configuration reactant liquor B, difference is that the consumption of ethylene glycol dimethacrylate is 94 μ L;
5) the resulting surface coverage of step 2 there are 4 layers of chip of unimolecule alkyl chain layer 4 be fixed on the flow cell of surface plasma resonance sensor, the another side of flow cell covers with the BK7 glass sheet of saturating ultraviolet light, at the BK7 glass sheet UV-LED pointolite that wavelength is 365nm is installed behind, in flow cell, inject reactant liquor A, unimolecule alkyl chain layer 4 is fully contacted with polymerization liquid A, be the UV-LED pointolite irradiation polymerization liquid initiated polymerization of 365nm then with wavelength, simultaneously, monitor the variation of sensing chip polymer thin film thickness in real time with surface plasma resonance sensor, stop illumination behind the illumination 70min, make it have the molecular imprinted polymer membrane of template molecule at 4 layers of chip surface growth one decks, with 5 layers of chip of gained material called after;
6) with step 5 resulting polymerization liquid B is injected reaction tank, be the UV-LED pointolite irradiation polymerization liquid B initiated polymerization of 365nm with wavelength, simultaneously, monitor the variation of sensing chip polymer thin film thickness in real time with surface plasma resonance sensor, stop illumination behind the illumination 30min, make it continue to form at the resulting 5 layers of chip of step 5 and have second layer molecular engram film, obtain having 6 layers of chip of the molecular imprinted polymer membrane of template molecule;
7) distilled water is injected reaction tank, make the environment of distilled water in the resulting 6 layers of abundant adaptive response of the chip pond of step 6, and carry out dynamics with surface plasma resonance sensor and monitor, intensity variations in the observing response pond, behind the 40min, when reflective light intensity no longer changed along with the time, reaction stopped;
8) for the eluent of 9:1 6 layers of chip that have molecular engram film are carried out wash-out by volume with distilled water and acetic acid, remove the template molecule in the molecular engram film, obtain the described a kind of molecular engram sensing chip that can detect testosterone at aqueous phase of present embodiment, SPR curve before and after the scanning wash-out, as shown in Figure 2;
Can be got by Fig. 2: the resonance angle of SPR curve is 69.4 degree before the wash-out, resonance angle 68.95 degree of curve behind the wash-out, and resonance angle has reduced by 0.45 degree, proves that eluent can wash away the template molecule on the molecular engram film.
9) with the aqueous solution of the sensing chip detection testosterone for preparing gained, solution concentration is 10
-12Mol/L, the SPR curve as shown in Figure 3 before and after the resulting absorption.
Can be got by Fig. 3: when the aqueous solution that contains sample was passed through flow cell, SPR curve resonance angle had increased by 0.3 degree, and curve is subjected to displacement, and shows that the prepared molecular engram sensing chip of the present invention can have response to the sample in the water, can detect the sample in the water.
The new construction of a kind of molecular engram sensing chip of the present invention as shown in Figure 1, comprises glass substrate 1, chromium rete 2, golden membranous layer 3, unimolecule alkyl chain layer 4,1 layer 5 of molecular imprinted polymer membrane, 2 layer 6 of molecular imprinted polymer membrane; This chip is followed successively by glass substrate 1, chromium rete 2, golden membranous layer 3, unimolecule alkyl chain layer 4, first molecular imprinted polymer membrane 5, second molecular imprinted polymer membrane 6 from lower to upper;
Be template molecule with the progesterone, prepare a kind of molecular engram sensing chip with new construction with light-initiated method, its concrete steps are:
1) adopt the method for vacuum evaporation at glass substrate 1 plating one deck chromium film and one deck gold film; The method of concrete evaporation is: with the chromium that the glass substrate 1 that cleans carries out vacuum evaporation 99.99%, vacuum tightness is 〉=1 * 10
-5Pa, evaporation speed is
The thickness of chromium film is 3nm; The then gold of evaporation 99.99% on chromium film 2, the thickness of gold-plated film be 43nm; The chip that obtain this moment is 3 layers of chip;
Wherein, described glass substrate 1 is the lanthanum glass of 2.5cm * 30cm;
3 layers of chip that 2) will newly plate immerse the ethanolic solution of the lauryl mercaptan of 50mM immediately, soak 24h, soak the back self assembly one deck unimolecule alkyl chain layer 4 on golden membranous layer 3 that finishes, and it is used alcohol flushing, and it is standby that nitrogen dries up the back, obtains 4 layers of chip;
Wherein, described lauryl mercaptan solution is the ethanolic solution of 50mmol/L lauryl mercaptan, and chemical formula is SH (CH
2)
11CH
3
3) 0.014g progesterone and 17 μ L methacrylic acids are dissolved in the 2mL acetonitrile in, ultrasonic mixing 10min leaves standstill 3h; Add again 37.5 μ L ethylene glycol dimethacrylates and with 0.008g benzophenone, ultrasonic mixing 10min, logical nitrogen 10min then, preparation prepolymerization reaction liquid A;
4) with step 3 configuration reactant liquor B, difference is that the consumption of ethylene glycol dimethacrylate is 56.5 μ L;
5) the resulting 4 layers of chip of step 2 are fixed on the flow cell of surface plasma resonance sensor, the another side of flow cell covers with the BK7 glass sheet of saturating ultraviolet light, at the BK7 glass sheet UV-LED pointolite that wavelength is 365nm is installed behind, in flow cell, inject reactant liquor A, unimolecule alkyl chain layer 4 is fully contacted with polymerization liquid A, be the UV-LED pointolite irradiation polymerization liquid initiated polymerization of 365nm then with wavelength, simultaneously, monitor the variation of sensing chip polymer thin film thickness in real time with surface plasma resonance sensor, stop illumination behind the illumination 64min, make it have the molecular imprinted polymer membrane of template molecule at 4 layers of chip surface growth one decks, with 5 layers of chip of gained material called after;
6) with step 5 resulting polymerization liquid B is injected reaction tank, be the UV-LED pointolite irradiation polymerization liquid B initiated polymerization of 365nm with wavelength, simultaneously, monitor the variation of sensing chip polymer thin film thickness in real time with surface plasma resonance sensor, stop illumination behind the illumination 20min, make it continue to form at the resulting 5 layers of chip of step 5 and have second layer molecular engram film, obtain having 6 layers of chip of the molecular imprinted polymer membrane of template molecule;
7) distilled water is injected reaction tank, make the environment of distilled water in the resulting 6 layers of abundant adaptive response of the chip pond of step 6, and carry out dynamics with surface plasma resonance sensor and monitor, intensity variations in the observing response pond, behind the 50min, reflective light intensity no longer changes along with the time, and reaction stops;
8) for the eluent of 8:2 6 layers of chip that have molecular engram film are carried out wash-out by volume with distilled water and acetic acid, remove the template molecule in the molecular engram film, obtain the described a kind of molecular engram sensing chip that can detect progesterone at aqueous phase of present embodiment, SPR curve before and after the scanning wash-out, as shown in Figure 4;
Can be got by Fig. 4: the resonance angle of SPR curve is 68.35 degree before the wash-out, resonance angle 67.85 degree of curve behind the wash-out, and resonance angle has reduced by 0.5 degree, proves that eluent can wash away the progesterone molecule on the molecular engram film;
9) with the aqueous solution of the sensing chip detection progesterone for preparing gained, solution concentration is 10
-14Mol/L, the SPR curve is as shown in Figure 5 before and after the resulting absorption;
Can be got by Fig. 5: when the aqueous solution that contains sample was passed through flow cell, SPR curve resonance angle had increased by 0.35 degree, and curve is subjected to displacement, and shows that the prepared molecular engram sensing chip of the present invention can have response to the sample in the water, can detect the sample in the water.
A kind of molecular engram sensing chip that can be used for detecting in the water has sandwich construction; Formed by glass substrate 1, metal film, unimolecule alkyl chain layer 4, multilayer molecular blotting membrane; This chip is arranged in order according to glass substrate 1, metal film, unimolecule alkyl chain layer 4, multilayer molecular blotting membrane order from the bottom to top.
The preparation method of this sensing chip, concrete steps are as follows:
1) adopt the method for vacuum evaporation at glass substrate 1 plating one deck chromium film and one deck gold film; The method of concrete evaporation is: with the chromium that the glass substrate 1 that cleans carries out vacuum evaporation 99.99%, vacuum tightness is 〉=1 * 10
-5Pa, evaporation speed is
The thickness of chromium film is 2.5nm; The then gold of evaporation 99.99% on chromium film 2, the thickness of gold-plated film be 45nm; The chip that obtain this moment is 3 layers of chip;
Wherein, described glass substrate 1 is the lanthanum glass of 2.5cm * 30cm;
3 layers of chip that 2) will newly plate immerse the ethanolic solution of the lauryl mercaptan of 100mM immediately, soak 24h, soak the back self assembly one deck unimolecule alkyl chain layer 4 on golden membranous layer 3 that finishes, and it is used alcohol flushing, and it is standby that nitrogen dries up the back, obtains 4 layers of chip;
Wherein, described lauryl mercaptan solution is the ethanolic solution of 50mmol/L lauryl mercaptan, and chemical formula is SH (CH
2)
11CH
3
3) 0.021g estradiol and 25.5 μ L methacrylic acids are dissolved in the 3mL acetonitrile in, ultrasonic mixing 7min leaves standstill 3.5h; Add again 56 μ L ethylene glycol dimethacrylates and with 0.012g benzophenone, ultrasonic mixing 10min, logical nitrogen 10min then, preparation prepolymerization reaction liquid A;
4) with step 3 configuration reactant liquor B, difference is that the consumption of ethylene glycol dimethacrylate is 70 μ L;
5) with step 3 configuration reaction liquid C, difference is that the consumption of ethylene glycol dimethacrylate is 78 μ L;
6) with step 3 configuration reactant liquor D, difference is that the consumption of ethylene glycol dimethacrylate is 85 μ L;
7) the resulting 4 layers of chip of step 2 are fixed on the flow cell of surface plasma resonance sensor, the another side of flow cell covers with the BK7 glass sheet of saturating ultraviolet light, at the BK7 glass sheet UV-LED pointolite that wavelength is 365nm is installed behind, in flow cell, inject reactant liquor 1, unimolecule alkyl chain layer 4 is fully contacted with polymerization liquid 1, be the UV-LED pointolite irradiation polymerization liquid initiated polymerization of 365nm then with wavelength, simultaneously, monitor the variation of sensing chip polymer thin film thickness in real time with surface plasma resonance sensor, stop illumination behind the illumination 60min, make it have the molecular imprinted polymer membrane of template molecule at 4 layers of chip surface growth one decks, with 5 layers of chip of gained material called after;
8) with step 7, resulting polymerization liquid B is injected reaction tank, be the UV-LED pointolite irradiation polymerization liquid B initiated polymerization of 365nm with wavelength, simultaneously, monitor the variation of sensing chip polymer thin film thickness in real time with surface plasma resonance sensor, stop illumination behind the illumination 30min, make it continue to form at the resulting 5 layers of chip of step 7 and have second layer molecular engram film, obtain having 6 layers of chip of the molecular imprinted polymer membrane of template molecule;
9) with step 7, resulting polymerization liquid C is injected reaction tank, be the UV-LED pointolite irradiation polymerization liquid C initiated polymerization of 365nm with wavelength, simultaneously, monitor the variation of sensing chip polymer thin film thickness in real time with surface plasma resonance sensor, stop illumination behind the illumination 20min, make it continue to form at the resulting 6 layers of chip of step 8 and have the 3rd layer of molecular engram film, obtain having 7 layers of chip of the molecular imprinted polymer membrane of template molecule;
10) with step 7, resulting polymerization liquid D is injected reaction tank, be the UV-LED pointolite irradiation polymerization liquid D initiated polymerization of 365nm with wavelength, simultaneously, monitor the variation of sensing chip polymer thin film thickness in real time with surface plasma resonance sensor, stop illumination behind the illumination 15min, make it continue to form at the resulting 7 layers of chip of step 9 and have the 4th layer of molecular engram film, obtain having 8 layers of chip of the molecular imprinted polymer membrane of template molecule;
11) distilled water is injected reaction tank, make the environment of distilled water in the resulting 8 layers of abundant adaptive response of the chip pond of step 10, and carry out dynamics with surface plasma resonance sensor and monitor, intensity variations in the observing response pond, behind the 60min, reflective light intensity no longer changes along with the time, and reaction stops;
12) for the eluent of 7:3 8 layers of chip that have molecular engram film are carried out wash-out by volume with distilled water and acetic acid, remove the estradiol molecule in the molecular engram film, obtain the described a kind of molecular engram sensing chip that can detect estradiol at aqueous phase of present embodiment;
13) with the aqueous solution of the sensing chip detection estradiol for preparing gained, solution concentration is 10
-13Mol/L, sensing chip has response to solution.
Claims (9)
1. a molecular engram sensing chip that can be used for detecting in the water is characterized in that: have sandwich construction; Formed by glass substrate, metal film, unimolecule alkyl chain layer, multilayer molecular blotting membrane; This chip is arranged in order according to glass substrate, metal film, unimolecule alkyl chain layer, multilayer molecular blotting membrane order from the bottom to top.
2. a kind of molecular engram sensing chip that can be used for detecting in the water as claimed in claim 1 is characterized in that: the preparation method of this sensing chip, and concrete steps are as follows:
The method of step 1, employing vacuum evaporation is at glass substrate plating one deck chromium film; Follow evaporation one deck gold film; With the 3 layers of chip of material called after that obtain;
Step 2,3 layers of chip of step 1 gained are dipped in the alkyl sulfhydryl solution; Immersion finishes the back at film surface self assembly last layer unimolecule alkyl chain layer, obtains the chip that surface coverage has unimolecule alkyl chain layer, with 4 layers of chip of its called after;
Step 3, the function monomer that template molecule is corresponding with it be dissolved in the corresponding pore-foaming agent of template molecule in, ultrasonic mixing, leave standstill after; Add and the corresponding crosslinking chemical of template molecule and the initiating agent corresponding with initiation conditions, nitrogen is led in ultrasonic mixing then, obtains polymerization liquid A again;
Step 4, dispose multiple polymerization liquid according to demand, the same step 3 of layoutprocedure, different is the amount of substance of employed crosslinking chemical, crosslinking chemical outside the amount in the layers of polymer reactant liquor greater than the amount of internal layer polymerization liquid;
Step 5, the reactant liquor A that step 3 is obtained inject reaction tank; 4 layers of chip that obtain in the step 2 place reaction tank top, make 4 layers of chip have the one side of unimolecule alkyl chain layer and contact with reactant liquor in the reaction tank; The condition of initiation is provided, makes to form the first molecular engram rete on the unimolecule alkyl chain layer, with 5 layers of chip of gained material called after;
Step 6, prepare the quantity of solution according to step 4, continue to form the multilayer molecular blotting membranes at 5 layers of chip of step 5 gained; The synthetic same step 5 of membrane process; The resulting multiple polymerization liquid of step 4 is injected reaction tank respectively; The condition of initiation is provided, makes having first molecular engram rete continuation formation multilayer molecular blotting membrane, with gained material called after multilayer molecular trace rete chip;
Step 7, configuration eluent carry out wash-out to the multilayer chiop of step 7 gained, remove the template molecule in the molecular engram film, and obtaining can be at the molecular engram sensing chip of aqueous phase test sample.
3. a kind of molecular engram sensing chip that can be used for detecting in the water as claimed in claim 1, it is characterized in that: more stable when detecting in water for the multilayer molecular trace rete chip that makes gained, can before the step 7 wash-out, handle chip, make it adapt to aqueous environment; Disposal route is with distilled water or PBS solution injection reaction tank, to make the aqueous environment in the abundant adaptive response of the resulting multilayer molecular trace of the step 6 rete chip pond.
4. a kind of molecular engram sensing chip that can be used for detecting in the water as claimed in claim 1 is characterized in that: the number of plies of the molecule engram film that the described multilayer molecular trace of step 6 rete chip forms is 2～5.
5. a kind of molecular engram sensing chip that can be used for detecting in the water as claimed in claim 1 is characterized in that: 4 layers of chip described in the step 2 need dry up with alcohol flushing and with nitrogen before use.
6. a kind of molecular engram sensing chip that can be used for detecting in the water as claimed in claim 1, it is characterized in that: the described template molecule of step 3 and step 4 is sample to be detected; The mol ratio of template molecule, function monomer and crosslinking chemical is 1:4:3～15; The volumetric molar concentration of template molecule in pore-foaming agent is 0.01～0.1mol/L, and the concentration of function monomer in pore-foaming agent is 0.04～0.4mol/L, and the concentration of crosslinking chemical in pore-foaming agent is 0.03～1.5mol/L; The concentration of initiating agent in pore-foaming agent is 1～10mg/ml.
7. a kind of molecular engram sensing chip that can be used for detecting in the water as claimed in claim 1, it is characterized in that: the amount of substance concentration of solute alkyl sulfhydryl is 1-100mmol/L in the alkyl sulfhydryl solution described in the step 2, and solvent is a kind of in methyl alcohol, ethanol or the methenyl choloride.
8. a kind of molecular engram sensing chip that can be used for detecting in the water as claimed in claim 1 is characterized in that: the described initiation conditions of step 5 and step 6 is for heat causes, ultraviolet light causes or the electropolymerization method.
9. a kind of molecular engram sensing chip that can be used for detecting in the water as claimed in claim 1, it is characterized in that: eluent described in the step 7 is the mixed solution of distilled water and acetic acid, and mixed volume is than 9:1～7:3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310177929.7A CN103234940B (en) | 2013-05-14 | 2013-05-14 | Molecular imprinting sensing chip available for underwater detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310177929.7A CN103234940B (en) | 2013-05-14 | 2013-05-14 | Molecular imprinting sensing chip available for underwater detection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103234940A true CN103234940A (en) | 2013-08-07 |
| CN103234940B CN103234940B (en) | 2015-05-13 |
Family
ID=48882991
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310177929.7A Expired - Fee Related CN103234940B (en) | 2013-05-14 | 2013-05-14 | Molecular imprinting sensing chip available for underwater detection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103234940B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105954233A (en) * | 2016-06-17 | 2016-09-21 | 北京理工大学 | Preparation method of surface plasma resonance sensor chip for detecting D-glucose in aqueous phase |
| CN106124457A (en) * | 2016-06-17 | 2016-11-16 | 北京理工大学 | A kind of preparation method of surface plasma resonance sensor chip based on nano material |
| CN106290565A (en) * | 2016-08-30 | 2017-01-04 | 上海交通大学 | Clenbuterol method in quickly detection pig urcine based on molecular engram film |
| CN107389619A (en) * | 2017-06-26 | 2017-11-24 | 北京理工大学 | A kind of surface plasma resonance sensor chip of aqueous phase detection estradiol and preparation method thereof |
| CN107941756A (en) * | 2017-11-15 | 2018-04-20 | 北京理工大学 | A kind of preparation method of hydrophilic surface plasma resonance sensing chip |
| CN113311049A (en) * | 2021-05-28 | 2021-08-27 | 江南大学 | Preparation method and application of molecular imprinting screen printing electrochemical sensor of progesterone molecules |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6525154B1 (en) * | 2000-07-20 | 2003-02-25 | The Regents Of The University Of California | Molecular imprinting for the recognition of peptides in aqueous solution |
| CN101324540A (en) * | 2008-07-17 | 2008-12-17 | 济南大学 | Trace Environmental Endocrine Disruptor Molecularly Imprinted Membrane Substrate and Its Preparation Method and Application |
| CN101571480A (en) * | 2009-06-08 | 2009-11-04 | 北京理工大学 | Surface plasma resonance detection method of morphine |
| CN102735647A (en) * | 2012-06-26 | 2012-10-17 | 北京理工大学 | Sensor chip, preparation method and use thereof |
-
2013
- 2013-05-14 CN CN201310177929.7A patent/CN103234940B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6525154B1 (en) * | 2000-07-20 | 2003-02-25 | The Regents Of The University Of California | Molecular imprinting for the recognition of peptides in aqueous solution |
| CN101324540A (en) * | 2008-07-17 | 2008-12-17 | 济南大学 | Trace Environmental Endocrine Disruptor Molecularly Imprinted Membrane Substrate and Its Preparation Method and Application |
| CN101571480A (en) * | 2009-06-08 | 2009-11-04 | 北京理工大学 | Surface plasma resonance detection method of morphine |
| CN102735647A (en) * | 2012-06-26 | 2012-10-17 | 北京理工大学 | Sensor chip, preparation method and use thereof |
Non-Patent Citations (1)
| Title |
|---|
| FENG SHI 等: "Surface Imprinting in Layer-by-Layer Nanostructured Films", 《ADV. FUNCT. MATER》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105954233A (en) * | 2016-06-17 | 2016-09-21 | 北京理工大学 | Preparation method of surface plasma resonance sensor chip for detecting D-glucose in aqueous phase |
| CN106124457A (en) * | 2016-06-17 | 2016-11-16 | 北京理工大学 | A kind of preparation method of surface plasma resonance sensor chip based on nano material |
| CN105954233B (en) * | 2016-06-17 | 2018-08-24 | 北京理工大学 | The preparation method of the surface plasma resonance sensor chip of detection D-Glucose in a kind of water phase |
| CN106124457B (en) * | 2016-06-17 | 2019-03-19 | 北京理工大学 | A kind of preparation method of the surface plasma resonance sensor chip based on nano material |
| CN106290565A (en) * | 2016-08-30 | 2017-01-04 | 上海交通大学 | Clenbuterol method in quickly detection pig urcine based on molecular engram film |
| CN106290565B (en) * | 2016-08-30 | 2019-07-12 | 上海交通大学 | Clenbuterol method in quick detection pig urcine based on molecular engram film |
| CN107389619A (en) * | 2017-06-26 | 2017-11-24 | 北京理工大学 | A kind of surface plasma resonance sensor chip of aqueous phase detection estradiol and preparation method thereof |
| CN107389619B (en) * | 2017-06-26 | 2020-02-14 | 北京理工大学 | Surface plasma resonance sensor chip for detecting estradiol in water phase and preparation method thereof |
| CN107941756A (en) * | 2017-11-15 | 2018-04-20 | 北京理工大学 | A kind of preparation method of hydrophilic surface plasma resonance sensing chip |
| CN113311049A (en) * | 2021-05-28 | 2021-08-27 | 江南大学 | Preparation method and application of molecular imprinting screen printing electrochemical sensor of progesterone molecules |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103234940B (en) | 2015-05-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103234940A (en) | Molecular imprinting sensing chip available for underwater detection | |
| Qi et al. | Three-dimensional paper-based microfluidic chip device for multiplexed fluorescence detection of Cu2+ and Hg2+ ions based on ion imprinting technology | |
| CN101887017A (en) | A surface plasmon resonance sensor chip and its preparation method | |
| CN101339128B (en) | Preparation method of surface plasma resonance imaging nano-structure array chip | |
| Wang et al. | Progress in the studies of photoelectrochemical sensors | |
| Zhang et al. | Surface plasmon resonance sensor for femtomolar detection of testosterone with water-compatible macroporous molecularly imprinted film | |
| Bao et al. | Detection of TNT by a molecularly imprinted polymer film-based surface plasmon resonance sensor | |
| CN102735647A (en) | Sensor chip, preparation method and use thereof | |
| CN106124457B (en) | A kind of preparation method of the surface plasma resonance sensor chip based on nano material | |
| CN102539388B (en) | A preparation method and application of a sensor chip for detecting atrazine | |
| Ko et al. | Porous silicon platform for optical detection of functionalized magnetic particles biosensing | |
| Janmanee et al. | Detection of human IgG on poly (pyrrole-3-carboxylic acid) thin film by electrochemical-surface plasmon resonance spectroscopy | |
| CN101498716A (en) | Methylamphetamine molecular recognition sensitive chip of sensor and its production method | |
| CN101551334B (en) | An optical sensor drug morphine molecular recognition sensitive chip and its preparation method | |
| Chah et al. | Ionization and reprotonation of self-assembled mercaptopropionic acid monolayers investigated by surface plasmon resonance measurements | |
| Decorbie et al. | Sensing polymer/paracetamol interaction with an independent component analysis-based SERS-MIP nanosensor | |
| CN101887016B (en) | A method for detecting testosterone using a surface plasmon resonance sensor | |
| CN105954233B (en) | The preparation method of the surface plasma resonance sensor chip of detection D-Glucose in a kind of water phase | |
| CN109507422B (en) | Optical microfluidic chips based on polymer and multi-layer metal nanoparticles decoration | |
| Ma et al. | A biomimetic mussel-inspired photoelectrochemical biosensing chip for the sensitive detection of CD146 | |
| Morita et al. | Surface plasmon resonance for detecting clenbuterol: Influence of monolayer structure | |
| JPH0627025A (en) | Molecule recognizing function film and sensor employing it | |
| Björk et al. | Dynamics of complex formation between biological and luminescent conjugated polyelectrolytes—A surface plasmon resonance study | |
| Shrestha et al. | Application of printable antibody ink for solid-phase immobilization of ABO antibody using photoactive hydrogel for surface plasmon resonance imaging | |
| CN102297851A (en) | Intelligent and hypersensitive molecular imprinting sensor chip |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CB02 | Change of applicant information |
Inventor after: Zhang Peng Inventor after: Han Shoupeng Inventor after: Meng Xiangyao Inventor after: Wei Tianxin Inventor after: Tan Yuan Inventor before: Wei Tianxin Inventor before: Tan Yuan |
|
| CB03 | Change of inventor or designer information | ||
| CP03 | Change of name, title or address | ||
| CP03 | Change of name, title or address |
Address after: 100161 Beijing City, Fengtai District Liuliqiao, No. 4 hospital Patentee after: The Chinese People's Liberation Army 92232 Troops Patentee after: BEIJING INSTITUTE OF TECHNOLOGY Address before: 100081 No. 5, Zhongguancun South Street, Haidian District, Beijing Patentee before: BEIJING INSTITUTE OF TECHNOLOGY |
|
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20150513 Termination date: 20200514 |