CN103234940A - Molecular imprinting sensing chip available for underwater detection - Google Patents
Molecular imprinting sensing chip available for underwater detection Download PDFInfo
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Abstract
The invention relates to a molecular imprinting sensing chip available for underwater detection and particularly relates to the field of molecular imprinting film sensing chips. The molecular imprinting sensing chip is in a multilayer structure and is composed of a glass substrate, a metal film, a monomolecular alkyl chain layer and a plurality layers of molecular imprinting films; and the glass substrate, the metal film, the monomolecular alkyl chain layer and the plurality layers of molecular imprinting films of the chip are sequentially arranged from bottom to top. An SPR (surface plasma resonance) sensor chip is prepared by a method of forming a plurality layer of molecular imprinting polymer films, thereby effectively weakening the destructive effect of water on hydrogen bonds in the molecular imprinting film to enable the synthetic molecular imprinting film to survive in water and sensitively detect template molecules in the water phase so as to provide a forceful detecting measure for accurately detecting the content of a sample in an actual environment. Therefore, the problem that a traditional chip is incapable of detecting the sample in the water phase can be effectively solved.
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.
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| 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 |
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| 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 |
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