CN102297851A - Intelligent and hypersensitive molecular imprinting sensor chip - Google Patents
Intelligent and hypersensitive molecular imprinting sensor chip Download PDFInfo
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- CN102297851A CN102297851A CN201110134911XA CN201110134911A CN102297851A CN 102297851 A CN102297851 A CN 102297851A CN 201110134911X A CN201110134911X A CN 201110134911XA CN 201110134911 A CN201110134911 A CN 201110134911A CN 102297851 A CN102297851 A CN 102297851A
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Abstract
The invention relates to an intelligent and hypersensitive molecular imprinting sensor chip, and specifically relates to the field of imprinting film preparation. The chip of the invention comprises a glass substrate, a metallic film and a molecular imprinting film arranged from bottom to top in order. The chip is prepared by the steps of: first dissolving a template molecule and functional monomer in a pore forming agent, and conducting ultrasonic mixing; adding a cross-linking agent and an initiator, and introducing nitrogen after ultrasonic mixing, thus obtaining a reacting solution; contacting the side of metallic film on the glass substrate plated with the metallic film with the reacting solution so as to initiate a polymerization reaction, thus forming the molecular imprinting film on the metallic film; and finally eluting the glass substrate so as to obtain the target product. With high detection sensitivity, the sensor chip of the invention can respond to a sample of ultra-low concentration. With strong specific recognition and wide scope of application, the sensor chip provided in the invention can be used in SPR (surface plasmon resonance) detection, QCM (quartz crystal microbalance) detection, SAW (surface acoustic wave) detection and electrochemical detection.
Description
Technical field
The present invention relates to the high sensitive molecule trace of a kind of intelligence sensing chip, be specifically related to the molecule engram film preparation field.
Background technology
(Molecular imprinting Technique MIT) is the cross discipline technology in conjunction with biological chemistry and high polymer chemistry that developed recently gets up to molecular imprinting.It utilize molecular engram film (Molecularimprinting Polymers, MIPs) in the simulation biology identification of antibody-antigentic specificity between interaction, (Template Molecule) carries out specific recognition to template molecule.The preparation molecular engram film generally includes three processes: (1) function monomer combines with the functional group of template molecule is reversible under suitable condition, forms host-guest complex; (2) add crosslinking chemical and initiating agent, under hot or light-initiated, make crosslinking chemical and function monomer reactive polymeric, in the polymkeric substance of formation is embedded in template molecule; (3) with certain physical and chemical method,, on the skeleton of polymkeric substance, stayed with the template molecule tool and " be scheduled to " optionally space and binding site template molecule wash-out from polymkeric substance.Owing to use each molecularly imprinted polymer of different template molecule systems to have different structures and character, so a kind of imprinted polymer can only combine with a kind of molecule, be similar to " lock " and " key ", imprinted polymer has narrow spectrum selection combination to this molecule in other words.
The molecular engram sensing chip is the core component of MIPs sensor, is one of molecular imprinting important application direction in recent years, is being widely used 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 preparation process of molecular engram film, the usage ratio of template molecule, function monomer and crosslinking chemical has tangible influence to polymer properties.The function monomer amount is very few, then can not take place to interact fully with template molecule; Otherwise, the self-polymerization of monomer then takes place easily, produce non-specific adsorption.The molar ratio of the crosslinking chemical that the tradition molecular engram film adopts is bigger, generally will account for the 70%-90% of raw material total amount, and wherein the mol ratio between template molecule, function monomer, the crosslinking chemical is 1: 4: 20～30.The molecular engram film of high crosslinking chemical ratio preparation is rigidity, good stability, but the sensitivity of rigid film is then relatively poor, can not produce response to the super low concentration sample.As Piletsky etc. the molecular engram film of atrazine being made conductivity sensor, only is 0.01-0.05mg/L to the sensing range of the atrazine in the solution.With phenylacrylic acid, sialic acid is the conductivity sensor of template molecule preparation, only can detect 10
-6The sample of mol/L concentration.
Summary of the invention
The objective of the invention is influences the problem of detection sensitivity in order to solve in the existing molecular engram sensing chip strong rigid molecule blotting membrane, proposes the high sensitive molecule trace of a kind of intelligence sensing chip and preparation method thereof.
The objective of the invention is to be achieved through the following technical solutions:
The high sensitive molecule trace of a kind of intelligence of the present invention sensing chip comprises glass substrate, metal film, molecular engram film, and this chip is arranged in order from the bottom to top according to the order of glass substrate, metal film, molecular engram film.
Preparation method's concrete steps of this sensing chip are:
1. the cleaning glass substrate dries up with nitrogen, plates layer of metal film with the method for vacuum evaporation on glass substrate, and thickness of metal film is 40～50nm;
2. under the lucifuge condition, template molecule, the function monomer corresponding with it be dissolved in the corresponding pore-foaming agent of template molecule in, ultrasonic mixing 5～40 minutes was left standstill 1～2 hour; Add crosslinking chemical corresponding and the initiating agent corresponding again with initiation conditions with template molecule, ultrasonic mixing 5～20 minutes, logical then nitrogen 5～30 minutes obtains reactant liquor;
3. the reactant liquor that step 2 is obtained injects reaction tank, the glass substrate of the metal-plated membrane that obtains in the step 1 is added on the reaction tank, its metal film one side is contacted with reactant liquor in the reaction tank, the condition of initiated polymerization is provided, make and form molecular engram film on the metal film, reaction time, the reaction time was long more by required thickness decision, and the molecular engram film of producing is thick more;
4. use pore-foaming agent corresponding and acetate preparation in 9: 1 by volume～7: 3 eluent with template molecule, the glass substrate that has molecular engram film that step 3 is obtained carries out wash-out, remove the template molecule in the molecular engram film, molecular engram film behind the wash-out is stayed have the same space configuration and contain the hole in selective binding site, obtain intelligent high sensitive molecule trace sensing chip with template molecule.
Wherein the described template molecule of step 2 is a sample molecule to be detected; Template molecule: function monomer: the mol ratio of crosslinking chemical is 1: 4: 3～15; The 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; The concentration of crosslinking chemical in pore-foaming agent is 0.03～1.5mol/L, and the concentration of initiating agent in pore-foaming agent is 1～10mg/ml;
The described initiation conditions of step 3 is heat initiation or light-initiated, and the initiating agent difference that different initiation conditions are used adopts azoisobutyronitrile as initiating agent when causing as heat, use benzophenone to be initiating agent when light-initiated.
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, after adding crosslinking chemical and initiating agent and initiated polymerization, on metallic film surface, grow one deck and contain the molecular engram film of template molecule, carry out wash-out with eluent then, eluent is by pore-foaming agent and acetate 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, stay the hole that has the same space structure with template molecule and contain the selective binding site, this cavity energy specific recognition and seizure sample molecule cause the variations in refractive index at interface between molecular engram film and the metal film.
Behind the adsorption sample molecule, molecular engram film contains the zone of convergency of discerning the hole and template molecule and interacts and cause the variation of surface charge, the polymer backbone conformation is recombinated, because the usage ratio of crosslinking chemical is very little in this chip, it is flexible that the molecular engram film of preparation is, the polymer backbone conformation change of molecular engram film is bigger, has increased the variations in refractive index at interface between molecular engram film and the metal film.
Beneficial effect
The high sensitive molecule trace of intelligence of the present invention sensing chip, can improve the detection sensitivity of SPR chip, make this chip produce response to the super low concentration sample, specific recognition is strong, applied widely, can be used for SPR detection, QCM detection, SAW detection and Electrochemical Detection, the molecular engram film thickness of chip surface can carry out accuracy controlling by reaction time length, the preparation nanoscale is to the film of micron order dimensional thickness, adopt in-situ polymerization technology, avoided pulverizing, later stage operation such as fixing, production cost is low, helps 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 map of the sensing chip that obtains among the embodiment 1;
Fig. 3 detects the SPR curve map of testosterone element for adopting the sensing chip that obtains among the embodiment 1;
Fig. 4 is the SPR curve map of the sensing chip that obtains among the embodiment 2;
Fig. 5 detects the SPR curve map of atrazine for adopting the sensing chip that obtains among the embodiment 2;
Among the figure, 1-glass substrate, 2-gold film, 3-molecular engram rete
Embodiment
With the application of the high sensitive molecule trace of intelligence of the present invention sensing chip in the SPR detector, in conjunction with the accompanying drawings and embodiments content of the present invention is described further below.
Embodiment 1
The high sensitive molecule trace of a kind of intelligence of the present invention sensing chip as shown in Figure 1, comprises glass substrate 1, golden film 2, and molecular engram film 3, this chip is arranged in order from the bottom to top according to the order of glass substrate, metal film, molecular engram film;
With the testosterone element is the method that template molecule prepares intelligent high sensitive molecule trace sensing chip, and its concrete steps are:
1. the cleaning glass substrate dries up with nitrogen, plates one deck gold film with the method for vacuum evaporation on substrate, and evaporation rate is 1
/ S, golden film thickness are 43nm;
2. in brown vial, add template molecule testosterone element, function monomer methacrylic acid and pore-foaming agent acetonitrile, testosterone element and methacrylic acid concentration in acetonitrile are respectively 0.025mol/L and 0.15mol/L, ultrasonic mixing 20 minutes, left standstill 2 hours, add crosslinking chemical ethylene glycol dimethacrylate and initiating agent azoisobutyronitrile then, ethylene glycol dimethacrylate and azoisobutyronitrile concentration in acetonitrile is respectively 0.2mol/L and 1.5mg/ml, ultrasonic mixing 10 minutes fed nitrogen 5 minutes;
3. the reaction solution with preparation in the step 2 adds in the reaction tank, covers the substrate that is coated with golden film that step 1 obtains then on reaction tank, and the one side of its golden film is contacted with reactant liquor, reacts 6 hours down at 60 ℃, obtains nano level molecular engram film;
4. prepare eluent with pore-foaming agent acetonitrile and acetate in 9: 1 ratio, the glass substrate that has molecular engram film to gained in the step 3 carries out wash-out, remove the plain molecule of testosterone on the molecular engram film, elution time is 30 minutes, make the hole that obtains having the same space configuration on the molecular engram film behind the wash-out and contain the selective binding site with template molecule, obtain molecular engram sensing chip based on the high sensitive molecule blotting membrane of flexibility, the SPR scanning curve of gained as shown in Figure 2, the resonance angle of molecular engram sensing chip is 58.2 ° before the polyreaction, and the resonance angle of molecular engram sensing chip is 72.5 ° after the polyreaction;
5. detect the acetonitrile solution of testosterone element with the sensing chip of preparation gained, solution concentration is 10
-12Mol/L, gained SPR curve data as shown in Figure 3.
The high sensitive molecule trace of a kind of intelligence of the present invention sensing chip comprises glass substrate 1, golden film 2, and molecular engram film 3, this chip is arranged in order from the bottom to top according to the order of glass substrate, metal film, molecular engram film;
With the atrazine is the method that template molecule prepares intelligent high sensitive molecule trace sensing chip, and its concrete steps are:
1. the cleaning glass substrate dries up with nitrogen, plates one deck gold film with the method for vacuum evaporation on substrate, and evaporation rate is 1
/ S, golden film thickness are 43nm;
2. in brown vial, add template molecule atrazine, function monomer methacrylic acid and acetonitrile, atrazine and methacrylic acid concentration in acetonitrile is respectively 0.025mol/L and 0.075mol/L, ultrasonic mixing 5 minutes, left standstill 1 hour, add crosslinking chemical ethylene glycol dimethacrylate and initiating agent benzophenone then, ethylene glycol dimethacrylate and benzophenone concentration in acetonitrile is respectively 0.1mol/L and 4mg/ml, and ultrasonic mixing 5 minutes fed nitrogen 5 minutes;
3. the reaction solution with preparation in the step 2 adds in the reaction tank, covers the substrate that is coated with golden film described in the step 1 then on reaction tank, and the one side of its golden film is contacted with reactant liquor, with UV rayed reaction 2 hours, obtains nano level molecular engram film;
4. prepare eluent with pore-foaming agent and acetate in 7: 3 ratio, the glass substrate that has molecular engram film that step 3 is obtained carries out wash-out, remove the atrazine molecule on the molecular engram film, elution time is 30 minutes, make the hole that obtains having the same space configuration on the molecular engram film behind the wash-out and contain the selective binding site with template molecule, obtain molecular engram sensing chip based on the high sensitive molecule blotting membrane of flexibility, the SPR scanning curve of gained chip as shown in Figure 4, the resonance angle of molecular engram sensing chip is 58.2 ° before the polyreaction, and the resonance angle of molecular engram sensing chip is 72.5 ° after the polyreaction;
5. detect the acetonitrile solution of atrazine with the sensing chip of preparation gained, solution concentration is 10
-16Mol/L, gained SPR curve data as shown in Figure 5.
Claims (1)
1. the high sensitive molecule trace of intelligence sensing chip, comprise glass substrate, metal film, molecular engram film, this chip is arranged in order from the bottom to top according to the order of glass substrate, metal film, molecular engram film, it is characterized in that preparation method's concrete steps of this sensing chip are:
1) clean glass substrate, dry up with nitrogen, plate layer of metal film with the method for vacuum evaporation on glass substrate, thickness of metal film is 40～50nm;
2) under the lucifuge condition, template molecule, the function monomer corresponding with it be dissolved in the corresponding pore-foaming agent of template molecule in, ultrasonic mixing 5～40 minutes was left standstill 1～2 hour; Add crosslinking chemical corresponding and the initiating agent corresponding again with initiation conditions with template molecule, ultrasonic mixing 5～20 minutes, logical then nitrogen 5～30 minutes obtains reactant liquor;
3) reactant liquor that step 2 is obtained injects reaction tank, the glass substrate of the metal-plated membrane that obtains in the step 1 is added on the reaction tank, its metal film one side is contacted with reactant liquor in the reaction tank, the condition of initiated polymerization is provided, make and form molecular engram film on the metal film;
4) use pore-foaming agent corresponding and acetate preparation in 9: 1 by volume～7: 3 eluent with template molecule, the glass substrate that has molecular engram film that step 3 is obtained carries out wash-out, remove the template molecule in the molecular engram film, obtain intelligent high sensitive molecule trace sensing chip;
Wherein the described template molecule of step 2 is a sample molecule to be detected; Template molecule: function monomer: the mol ratio of crosslinking chemical is 1: 4: 3～15; The 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; The concentration of crosslinking chemical in pore-foaming agent is 0.03～1.5mol/L, and the concentration of initiating agent in pore-foaming agent is 1～10mg/ml;
The described initiation conditions of step 3 is heat initiation or light-initiated.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103698242A (en) * | 2013-12-21 | 2014-04-02 | 中国科学院苏州生物医学工程技术研究所 | Rapid detecting sensor for microcystins |
| 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 |
| CN109073561A (en) * | 2016-01-29 | 2018-12-21 | 梁庆耀 | Detection of Organic Chemicals |
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| WO2001077664A2 (en) * | 2000-04-10 | 2001-10-18 | The Johns Hopkins University | Molecularly imprinted polymeric sensor for the detection of explosives |
| US20040072373A1 (en) * | 2002-10-15 | 2004-04-15 | Industrial Technology Research Institute | Biochemical labeling materials and manufacturing method thereof |
| CN101498716A (en) * | 2009-03-11 | 2009-08-05 | 北京理工大学 | Methylamphetamine molecular recognition sensitive chip of sensor and its production method |
| CN101551334B (en) * | 2009-03-05 | 2011-02-09 | 公安部物证鉴定中心 | Drug morphia molecular recognition sensitivity chip of optical sensor and producing method thereof |
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2011
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2001077664A2 (en) * | 2000-04-10 | 2001-10-18 | The Johns Hopkins University | Molecularly imprinted polymeric sensor for the detection of explosives |
| US20040072373A1 (en) * | 2002-10-15 | 2004-04-15 | Industrial Technology Research Institute | Biochemical labeling materials and manufacturing method thereof |
| CN101551334B (en) * | 2009-03-05 | 2011-02-09 | 公安部物证鉴定中心 | Drug morphia molecular recognition sensitivity chip of optical sensor and producing method thereof |
| CN101498716A (en) * | 2009-03-11 | 2009-08-05 | 北京理工大学 | Methylamphetamine molecular recognition sensitive chip of sensor and its production method |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103698242A (en) * | 2013-12-21 | 2014-04-02 | 中国科学院苏州生物医学工程技术研究所 | Rapid detecting sensor for microcystins |
| CN103698242B (en) * | 2013-12-21 | 2016-09-28 | 中国科学院苏州生物医学工程技术研究所 | A kind of quick-check sensor of Microcystin |
| CN109073561A (en) * | 2016-01-29 | 2018-12-21 | 梁庆耀 | Detection of Organic Chemicals |
| 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 |
| 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 |
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Application publication date: 20111228 |