WO2014075482A1 - 一种表面等离子体共振传感芯片及其制备方法、应用 - Google Patents
一种表面等离子体共振传感芯片及其制备方法、应用 Download PDFInfo
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- WO2014075482A1 WO2014075482A1 PCT/CN2013/082150 CN2013082150W WO2014075482A1 WO 2014075482 A1 WO2014075482 A1 WO 2014075482A1 CN 2013082150 W CN2013082150 W CN 2013082150W WO 2014075482 A1 WO2014075482 A1 WO 2014075482A1
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- Prior art keywords
- glass substrate
- plasmon resonance
- surface plasmon
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- sensing chip
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- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 title claims abstract description 85
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/55—Specular reflectivity
- G01N21/552—Attenuated total reflection
- G01N21/553—Attenuated total reflection and using surface plasmons
-
- C—CHEMISTRY; METALLURGY
- C03—GLASS; MINERAL OR SLAG WOOL
- C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
- C03C17/00—Surface treatment of glass, not in the form of fibres or filaments, by coating
- C03C17/34—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions
- C03C17/36—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions at least one coating being a metal
- C03C17/38—Surface treatment of glass, not in the form of fibres or filaments, by coating with at least two coatings having different compositions at least one coating being a metal at least one coating being a coating of an organic material
-
- C—CHEMISTRY; METALLURGY
- C23—COATING METALLIC MATERIAL; COATING MATERIAL WITH METALLIC MATERIAL; CHEMICAL SURFACE TREATMENT; DIFFUSION TREATMENT OF METALLIC MATERIAL; COATING BY VACUUM EVAPORATION, BY SPUTTERING, BY ION IMPLANTATION OR BY CHEMICAL VAPOUR DEPOSITION, IN GENERAL; INHIBITING CORROSION OF METALLIC MATERIAL OR INCRUSTATION IN GENERAL
- C23C—COATING METALLIC MATERIAL; COATING MATERIAL WITH METALLIC MATERIAL; SURFACE TREATMENT OF METALLIC MATERIAL BY DIFFUSION INTO THE SURFACE, BY CHEMICAL CONVERSION OR SUBSTITUTION; COATING BY VACUUM EVAPORATION, BY SPUTTERING, BY ION IMPLANTATION OR BY CHEMICAL VAPOUR DEPOSITION, IN GENERAL
- C23C14/00—Coating by vacuum evaporation, by sputtering or by ion implantation of the coating forming material
- C23C14/06—Coating by vacuum evaporation, by sputtering or by ion implantation of the coating forming material characterised by the coating material
- C23C14/14—Metallic material, boron or silicon
- C23C14/18—Metallic material, boron or silicon on other inorganic substrates
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
-
- C—CHEMISTRY; METALLURGY
- C03—GLASS; MINERAL OR SLAG WOOL
- C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
- C03C17/00—Surface treatment of glass, not in the form of fibres or filaments, by coating
- C03C17/06—Surface treatment of glass, not in the form of fibres or filaments, by coating with metals
- C03C17/09—Surface treatment of glass, not in the form of fibres or filaments, by coating with metals by deposition from the vapour phase
-
- C—CHEMISTRY; METALLURGY
- C03—GLASS; MINERAL OR SLAG WOOL
- C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
- C03C2217/00—Coatings on glass
- C03C2217/20—Materials for coating a single layer on glass
- C03C2217/25—Metals
- C03C2217/251—Al, Cu, Mg or noble metals
- C03C2217/254—Noble metals
- C03C2217/255—Au
-
- C—CHEMISTRY; METALLURGY
- C03—GLASS; MINERAL OR SLAG WOOL
- C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
- C03C2218/00—Methods for coating glass
- C03C2218/10—Deposition methods
- C03C2218/15—Deposition methods from the vapour phase
- C03C2218/151—Deposition methods from the vapour phase by vacuum evaporation
-
- C—CHEMISTRY; METALLURGY
- C03—GLASS; MINERAL OR SLAG WOOL
- C03C—CHEMICAL COMPOSITION OF GLASSES, GLAZES OR VITREOUS ENAMELS; SURFACE TREATMENT OF GLASS; SURFACE TREATMENT OF FIBRES OR FILAMENTS MADE FROM GLASS, MINERALS OR SLAGS; JOINING GLASS TO GLASS OR OTHER MATERIALS
- C03C2218/00—Methods for coating glass
- C03C2218/10—Deposition methods
- C03C2218/15—Deposition methods from the vapour phase
- C03C2218/154—Deposition methods from the vapour phase by sputtering
- C03C2218/156—Deposition methods from the vapour phase by sputtering by magnetron sputtering
Definitions
- the invention belongs to the field of surface plasmon sensor chip preparation, and in particular relates to a surface plasmon resonance sensor chip which can be used for lipopolysaccharide detection, a preparation method thereof and an application thereof.
- Lipopolysaccharide is a polymer composed of a lipid and a polysaccharide linked by a covalent bond. It is a major component of the outer membrane of Gram-negative bacteria and is a potent bacterial toxin known as endotoxin. When gram-negative bacteria such as Escherichia coli and Salmonella enterica are multiplied or lysed, lipopolysaccharide is released. Gram-negative bacteria in the human body release a large amount of lipopolysaccharide in the cell wall, which causes inflammation and inflammation of the human body, and further develops into a serious disease that seriously threatens human health - sepsis.
- LPS Lipopolysaccharide
- the content of LPS must be detected in purified water and water for injection to ensure water quality.
- researchers have therefore been working to invent a highly selective, ultrasensitive method for the detection of lipopolysaccharides in aqueous solutions.
- the gel method is the most commonly used method for detecting lipopolysaccharide in clinical practice. It can detect lipopolysaccharide qualitatively, semi-quantitatively and highly sensitively, but this method must use a paleontological ( ⁇ ) blood, so Long-term and large-scale use must be subject to certain restrictions.
- SPR Surface Plasmon Resonance
- a total internal reflection evanescent wave is generated, which has a wave penetration distance of about 300 nm, which can induce surface electrons generated by free electrons on the metal surface.
- the surface plasmons are equal to the frequency of the evanescent wave, the two will resonate, and the incident light is absorbed, causing the reflected light energy to drop sharply, so that the resonance peak appears at the reflection spectrum (ie, the lowest value of the reflection intensity).
- any slight change in the refractive index and conformation of the surface medium will cause the angle of incidence to migrate, which is captured by the detector and converted into the corresponding spectrum. Since the surface plasma wave is very sensitive to the small change of the refractive index and conformation of the medium, if the sample to be tested is brought into contact with the metal film where surface plasmon resonance occurs, and the interaction occurs, the dielectric constant, refractive index and conformation of the film will occur. The change, and thus the resonance condition, causes the shift of the formant.
- the resonance principle of the SPR sensor determines that this sensor has obvious advantages compared with traditional biological detection methods or chemical detection methods, such as Real time, dynamic, and especially ultra-sensitive detection.
- SPR sensors have been widely used in many fields such as environmental sanitation, food safety, and disease diagnosis. It is mainly used to detect interactions between biomolecules and other substances and kinetic molecules.
- SPR sensors for detecting lipopolysaccharide have not appeared, and SPR technology has not been used to detect the content of lipopolysaccharide in aqueous solution, especially the content of lipopolysaccharide in water for injection, in real time, quickly, simply, quantitatively and ultrasensitively.
- a first technical problem to be solved by the present invention is to provide a surface plasmon resonance sensor chip.
- a second technical problem to be solved by the present invention is to provide a method of preparing a surface plasmon resonance sensor chip.
- the surface plasmon resonance spectroscopy technique generated on the surface of the gold film on the glass substrate is used to quickly, easily, quantitatively and ultrasensitively detect the content of lipopolysaccharide in the aqueous solution.
- a third technical problem to be solved by the present invention is to provide an application of a surface plasmon resonance sensor chip.
- the invention provides a surface plasmon resonance sensor chip, which comprises a glass substrate layer, a gold film layer and a probe molecule layer; a gold film layer is disposed on the glass substrate layer, and a probe molecule layer is disposed on the gold film layer;
- the probe molecule layer is a layer of one or more probe molecules having the following structure:
- X, Y, W are O, S, NR 5 or Si-R 6 R respectively Zi ? , Ri, R2, R, , , R6, R7 ' R8, R9 ' Rl0, Rll, Rl2, Rl3, Rl4,
- R 15 is a hydrogen atom; a thiol group of 1 to 18 carbons, a hydroxyl group, a decyl group, a carboxyl group, an amide, an acid anhydride, an alkenyl group, an alkynyl group, an aryl group, an ester group, an ether group, an amino group, a cyano group, a quaternary ammonium salt, Sulfonate, phosphate or polyethylene glycol; m, n, 0 is 0 to 10000, and is not 0 at the same time.
- “1 to 18 carbon sulfhydryl groups, hydroxy groups, mercapto groups, carboxyl groups, amides, acid anhydrides, alkenyl groups, alkynyl groups, aryl groups, ester groups, ether groups” means sulfhydryl groups, hydroxyl groups, sulfhydryl groups, carboxyl groups, amide groups.
- the carbon of the acid anhydride, alkenyl group, alkynyl group, aryl group, ester group or ether group is 1 to 18 carbons.
- the thickness of the gold film layer is 10 to 60 nm ; the thickness of the probe molecule layer is 1 to 100 nm ; and the coverage of the probe molecule layer on the gold film layer is 5% to 100%.
- the coverage of the probe molecule on the gold film layer can be semi-quantitatively determined by AFM; after the probe molecule covers the gold film layer, it will self-assemble to form a granular structure; the surface of the probe molecule is modified with a sulfhydryl group and a gold film. contact.
- the invention also provides a preparation method of a surface plasmon resonance sensor chip, comprising the following steps:
- the glass substrate obtained in the step 1) is completely immersed in a probe molecular solution having a concentration of 0.01 to 1000 mg/mL, and left for 5 minutes to 24 hours;
- the probe molecule solution is a solution having one or more of the following structures:
- Ar 2 is one of the following structures:
- X, Y, W are O, S, NR 5 or Si-R 6 R 7 respectively ;
- Ri , R 2 , R 3 , R4 , R5 , R 6 , R 7 , R 8 , Rg, R 10 , Ru, R 12 , R 13 , R 14 , R 15 are each a hydrogen atom; 1 ⁇ 18 Carbon sulfhydryl, hydroxy, decyl, carboxy, amide, anhydride, alkenyl, alkynyl, aryl, ester, ether, amino, cyano, quaternary ammonium, sulfonate, phosphate or polyethylene glycol
- the base; m, n, 0 is 0 ⁇ 10000, and is not 0 at the same time.
- the gold film has a thickness of 10 to 60 nm.
- the coverage of the probe molecule on the gold film is 5% to 100%.
- the waters that can be selected are: secondary distilled water, tertiary distilled water, four distilled water, ultrapure water, and the like.
- the rinsing solution After rinsing once or several times with water, the rinsing solution is analyzed with a high-performance liquid phase, and the signal of the probe molecule is not detected, and it can be considered that it has been rinsed clean.
- the purpose of the rinsing is to remove the probe molecules adsorbed on the surface of the gold film by physical absorption.
- the concentration of the probe molecules and the soaking time will ultimately affect the coverage of the probe molecules on the gold film, which in turn affects the sensitivity of the chip.
- the resulting sensor chip is stored in secondary distilled water for later use.
- the above reaction may be carried out at room temperature.
- Step 1) The coating is performed by vacuum evaporation or magnetron sputtering.
- the vacuum degree is 1 X 10 - 4 Pa
- the thickness of the gold film of 10 to 60 nm can be precisely controlled by adjusting the frequency variation of the film thickness meter (10 to 60 Hz) and the evaporation rate of 0. lA/s.
- the solvent of the probe molecule solution is physiological saline, HEPES buffer, phosphate buffer; or methanol, ethanol, acetonitrile, dichloromethane, trichloromethane, tetrahydrofuran, dimethyl sulfoxide, hydrazine, hydrazine - one or a mixture of two or more kinds of organic solvents such as dimethylformamide, hydrazine, hydrazine-dimethylacetamide; or a mixture of one or more of the foregoing organic solvents and water in any ratio Mixed mixture.
- the invention further provides a method for preparing a surface plasmon resonance sensor chip, comprising the following steps: (1) plating gold on the surface of a glass substrate;
- the glass substrate obtained in the step (1) is immersed in a solution of 0.01 to 1000 mmol/L of the compound S1, allowed to stand for 1 to 24 hours, rinsed repeatedly with water, and used; probe molecules, NHS, EDC, solvent According to the mass ratio of 1: 0.1 ⁇ 100: 0.1-100: 1-1000, a mixed solution is obtained; the glass substrate is immersed in the mixed solution, allowed to stand for 5 minutes to 24 hours, the glass substrate is taken out, and ethanol is used. The water is repeatedly washed; wherein, the structure of the compound S1 is:
- R 16 are each a hydrogen atom; a thiol group of 1 to 18 carbons, a hydroxyl group, a decyl group, a carboxyl group, an amide group, an acid anhydride group, an alkenyl group, an alkynyl group, an aryl group, an ester group, an ether group, an amino group, a cyano group Or a polyethylene glycol group;
- the structure of the probe molecule is defined as defined in the above paragraph.
- NHS is N-hydroxysuccinimide
- EDC is 1-ethyl-(3-dimethylaminopropyl) carbodiimide.
- the gold film has a thickness of 10 to 60 nm.
- the coverage of the probe molecule on the gold film is 5% to 100%.
- the waters that can be selected are: secondary distilled water, tertiary distilled water, four distilled water, ultrapure water, and the like.
- the signal of S1 is not detected, and it can be considered that it has been rinsed clean. Improper rinsing can affect the coverage of the probe molecules on the gold film.
- the soaked glass substrate was first rinsed with ethanol and then rinsed with water.
- the S1 compounds are either commercially available or synthesized by existing literature.
- the above reaction may be carried out at room temperature.
- Step (1) The coating is by vacuum evaporation or magnetron sputtering.
- the degree of vacuum is 1 X 10 - 4 Pa
- the thickness of the gold film of 10 to 60 nm can be precisely controlled by adjusting the frequency variation (10 to 60 Hz) of the film thickness meter and the evaporation rate of 0. lA/s.
- the compound S1 is dissolved in physiological saline, HEPES buffer, phosphate buffer; or methanol, ethanol, acetonitrile, dichloromethane, trichloromethane, tetrahydrofuran, dimethyl sulfoxide, hydrazine, hydrazine- One or a mixture of two or more kinds of organic solvents such as dimethylformamide, hydrazine, hydrazine-dimethylacetamide; or a mixture of one or more of the foregoing organic solvents and water in any ratio In the mixture, a solution of the compound S1 is formed.
- the solvent is physiological saline, HEPES buffer, phosphate buffer; or methanol,
- organic solvents such as ethanol, acetonitrile, methylene chloride, chloroform, tetrahydrofuran, dimethyl sulfoxide, hydrazine, hydrazine-dimethylformamide, hydrazine, hydrazine-dimethylacetamide
- the above mixed solvent or a mixture of one or a mixture of two or more of the foregoing organic solvents and water mixed in an arbitrary ratio.
- the invention also provides the use of a surface plasmon resonance sensor chip for detecting lipopolysaccharide in an aqueous solution.
- the application is to install the sensor chip into an angle modulation type or a wavelength modulation type surface plasmon resonance sensing device, and pass different concentrations of the lipopolysaccharide aqueous solution in the flow cell to detect the surface plasmon resonance peak
- the offset is used to detect lipopolysaccharide.
- the amount of shift of the surface plasmon resonance peak is linear with the concentration of the introduced lipopolysaccharide in a corresponding range, and can be used for quantitative detection of lipopolysaccharide.
- the present invention firstly proposes the use of surface plasmon resonance sensing technology to detect LPS in aqueous solution. Compared with the conventional detection method, the sensitivity is high and the selectivity is good. The linear range of detection of lipopolysaccharide concentration in aqueous solution is lO -lO ⁇ M;
- the modification of the probe molecule on the surface of the gold film used in the present invention has a clear structure, the molecule is easy to synthesize, and the structure is controllable;
- the process of preparing the SPR chip of the invention has the advantages of simple operation steps and low cost, so that the prepared chip has good reproducibility and meets the requirements of batch preparation in industrial production, so that it is easy to be practically popularized and applied.
- FIG. 1 is a schematic structural view of a sensor chip of the present invention.
- AFM atomic force microscope
- Fig. 3 is a diagram showing the selectivity test of the sensor chip of the present invention for lipopolysaccharide in an aqueous solution by a wavelength type surface plasmon resonance sensing device.
- the ordinate is a relative light intensity value at a wavelength shift of 12 nm of the resonance wavelength
- the abscissa is a lipopolysaccharide concentration
- Fig. 5 is a graph showing changes in the concentration of the sensor chip and lipopolysaccharide of Example 6.
- Fig. 6 is a graph showing changes in the concentration of the sensor chip and lipopolysaccharide of Example 15.
- Fig. 7 is a view showing the molecular structure of several probe molecules and compound S1 involved in the examples.
- a surface plasmon resonance sensor chip comprises a glass substrate layer, a gold film layer and a probe molecule layer; a gold film layer is disposed on the glass substrate layer, and a probe molecule layer is disposed on the gold film layer; the gold film layer a thickness of 10 ⁇ 60nm; the probe molecule layer thickness is l ⁇ 100nm; probe the molecular structure layer is a layer of one of the following structures or two or more probe molecules:
- X, Y, W are O, S, NR 5 or Si-R 6 R 7 respectively ;
- R 10 , Ru, R 12 , R 13 , R 14 , R 15 are each a hydrogen atom; 1 to 18 carbons of ruthenium Base, hydroxy, thiol, carboxy, amide, anhydride, alkenyl, alkynyl, aryl, ester, ether, amino, cyano, quaternary ammonium, sulfonate, phosphate or polyethylene glycol; m , n, 0 is 0 ⁇ 10000, and is not 0 at the same time.
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- Figure 2 is an atomic force microscope (AFM) image of a gold film plated on a glass substrate.
- AFM atomic force microscope
- the white particles in Figure 2 (C) are probe molecules with a coverage of approximately 20%.
- the surface plasmon resonance sensor chip prepared in Example 2 is incorporated into an angle modulation type or wavelength modulation type surface plasmon resonance sensing device, and different concentrations of the lipopolysaccharide aqueous solution are introduced into the flow cell to cause a reflection angle.
- the change by detecting the change in the angle, is linear with the concentration of the introduced lipopolysaccharide, and the results are shown in Fig. 3.
- the offset of the surface plasmon resonance peak is linear with the concentration of the introduced lipopolysaccharide in the range of 10-14 ⁇ 10 - 1Q M.
- the sensor chip is incorporated into a wavelength-type surface plasmon resonance sensing device, and a 10-1 Q M lipopolysaccharide aqueous solution is introduced into the flow cell, causing a large change in resonance wavelength and relative light intensity.
- an aqueous solution of other interfering substances of the same concentration such as sodium chloride, potassium chloride, calcium chloride, glucose, ATP, DNA, RNA, LPA (lysophosphatidic acid), SDS, LTA (lipophosphoic acid), etc. , causing a small change in the resonant wavelength and relative light intensity.
- the result is shown in Figure 4. It can be seen that the sensor chip of the present invention is selective for lipopolysaccharide.
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the gold film-coated glass substrate obtained in the step 1) is completely immersed in a probe molecule (PT1) solution having a concentration of 10 mg/mL, and the solvent is acetonitrile, and left at room temperature for 10 hours;
- PT1 probe molecule
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the gold film-coated glass substrate obtained in the step 1) is completely immersed in a probe molecule (PT1) having a concentration of 100 mg/mL, the solvent is 10% tetrahydrofuran and 90% water, and the temperature is 20 Hour
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- step 1) using a vacuum evaporation technique, plating a thickness of 50 nm gold on the surface of the glass substrate; 2)
- the gold film-coated glass substrate obtained in step 1) is completely immersed in a probe molecule (PT2) solution having a concentration of 1 mg/mL, and the solvent is 10% acetonitrile and 90% water, and left at room temperature. 3 hours;
- PT2 probe molecule
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the surface plasmon resonance sensor chip prepared in Example 6 is incorporated into a wavelength modulation type surface plasmon resonance sensing device, and different concentrations of the lipopolysaccharide aqueous solution are introduced into the flow cell, which causes a change in the resonance wavelength, and the detection is performed.
- the change in the resonant wavelength is linear with the concentration of the introduced lipopolysaccharide.
- the results are shown in Fig. 5.
- the offset of the surface plasmon resonance peak is linear with the concentration of the introduced lipopolysaccharide in the range of 10 - 1Q ⁇ 10 - 8 M.
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the gold film-coated glass substrate obtained in the step 1) is completely immersed in a probe molecule (PT2) solution having a concentration of 1 mg/mL, and the solvent is phosphate buffer solution water, and left at room temperature for 13 hours;
- PT2 probe molecule
- the glass substrate is taken out, and the glass substrate is repeatedly washed with ultrapure water to obtain a chip, which is stored in the secondary distilled water for use.
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the gold film-coated glass substrate obtained in the step 1) is completely immersed in a probe molecule (PT4) solution having a concentration of 15 mg/mL, and the solvent is physiological saline, and left at room temperature for 15 hours;
- PT4 probe molecule
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- step 1) 1) using a vacuum evaporation technique, plating a thickness of 60 nm on the surface of the glass substrate; 2) completely immersing the gold film-coated glass substrate obtained in step 1) in a probe molecule (PT4) solution having a concentration of 100 mg/mL, and the solvent is hydroxyethylpiperazine ethanesulfonic acid buffer solution, g ⁇ HEPE, leave it at room temperature for 5 hours;
- PT4 probe molecule
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the glass substrate obtained in the step 1) is immersed in a 0.01 mmol/L compound S1-1 solution, allowed to stand at room temperature for 1 hour; and repeatedly rinsed with double distilled water;
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the glass substrate obtained in the step 1) is immersed in a lmmol/L compound S1-2 solution, allowed to stand at room temperature for 4 hours; and repeatedly rinsed with double distilled water;
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the glass substrate obtained in the step 1) is immersed in a 10 mmol/L compound S1-3 solution, allowed to stand at room temperature for 10 hours; and repeatedly rinsed with double distilled water;
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the glass substrate obtained in the step 1) is immersed in a lmmol/L compound S1-4 solution, allowed to stand at room temperature for 10 hours; and repeatedly rinsed with double distilled water;
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the glass substrate obtained in the step 1) is immersed in a 100 mmol/L compound S1-5 solution, allowed to stand at room temperature for 10 hours; and repeatedly rinsed with double distilled water;
- the surface plasmon resonance sensor chip prepared in Example 15 is loaded into a wavelength modulation type surface plasmon resonance sensing device, and different concentrations of the lipopolysaccharide aqueous solution are introduced into the flow cell, which causes a change in the resonance wavelength, and the detection is performed.
- the change in the resonant wavelength is linear with the concentration of the introduced lipopolysaccharide.
- the results are shown in Fig. 6.
- the offset of the surface plasmon resonance peak is linear with the concentration of the introduced lipopolysaccharide in the range of 10-14 ⁇ 10 - 1Q M.
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the glass substrate obtained in the step 1) is immersed in a 0.1 mmol/L compound S1-6 solution, allowed to stand at room temperature for 24 hours; and repeatedly rinsed with double distilled water;
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the gold film-coated glass substrate obtained in the step 1) is completely immersed in a probe molecule PE1 solution having a concentration of 10 mg/mL, and the solvent is acetonitrile, and left at room temperature for 10 hours;
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the gold film-coated glass substrate obtained in the step 1) is completely immersed in a probe molecule PE2 solution having a concentration of 20 mg/mL, and the solvent is dimethyl sulfoxide, and left at room temperature for 10 hours;
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the gold film-coated glass substrate obtained in the step 1) is completely immersed in a probe molecule PPV2 solution having a concentration of 10 mg/mL, and the solvent is tetrahydrofuran, and allowed to stand at room temperature for 1 hour;
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the gold film-coated glass substrate obtained in the step 1) is completely immersed in a probe molecule PF solution having a concentration of 10 mg/mL, the solvent is water, and left at room temperature for 3 hours;
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the gold film-coated glass substrate obtained in the step 1) is completely immersed in a probe molecule PF2 solution having a concentration of 10 mg/mL, the solvent is water, and left at room temperature for 5 hours;
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- step 1) using a magnetron sputtering technique, plating a thickness of 48 nm gold on the surface of the glass substrate; 2)
- the gold film-coated glass substrate obtained in the step 1) is completely immersed in a probe molecule PPP1 solution having a concentration of 10 mg/mL, and the solvent is dichloromethane, and left at room temperature for 10 hours;
- a method for preparing a surface plasmon resonance sensor chip the specific steps are as follows:
- the gold film-coated glass substrate obtained in the step 1) is completely immersed in a probe molecule PPP2 solution having a concentration of 10 mg/mL, the solvent is water, and left at room temperature for 10 hours;
- a method for preparing a surface plasmon resonance sensor chip comprising the following steps:
- the glass substrate obtained in the step 1) is completely immersed in a probe molecule solution having a concentration of O.Omg/mL, and left for 5 minutes;
- the glass substrate was taken out, and the glass substrate was repeatedly washed with water to obtain a surface plasmon resonance sensor chip.
- the solvent of the probe molecule solution is hydrazine, hydrazine-dimethylformamide.
- the probe molecule is ⁇ 2.
- a method for preparing a surface plasmon resonance sensor chip comprising the following steps:
- the glass substrate obtained in the step 1) is completely immersed in a probe molecule solution having a concentration of 1000 mg/mL, and left for 24 hours;
- the glass substrate was taken out, and the glass substrate was repeatedly washed with water to obtain a surface plasmon resonance sensor chip.
- the solvent of the probe molecule solution is hydrazine, hydrazine-dimethylacetamide.
- the probe molecule is PF2.
- a method for preparing a surface plasmon resonance sensor chip comprising the following steps:
- the glass substrate obtained in the step (1) is immersed in the O.01 mmol/L compound S1 solution, allowed to stand for 1 hour, rinsed repeatedly with water, and used; the probe molecules, NHS, EDC, solvent are in accordance with the quality.
- the structure of the compound S1 is:
- Z 3 and R 16 are a hydrogen atom
- the solvent of the compound S1 solution is physiological saline.
- the solvent is HEPES buffer.
- a method for preparing a surface plasmon resonance sensor chip comprising the following steps:
- Gold is plated on the surface of the glass substrate, and the thickness of the gold film is 60 nm ;
- the glass substrate obtained in the step (1) is immersed in a 1000 mmol/L compound S1 solution, allowed to stand for 24 hours, rinsed repeatedly with water, and used; the probe molecule, NHS, EDC, solvent are mass-produced.
- 1 100: 100: 1000 mixed, to obtain a mixed solution; the aforementioned glass substrate was immersed in the mixed solution, allowed to stand for 24 hours, the glass substrate was taken out, and repeatedly washed with ethanol and water;
- the structure of the compound S1 is:
- R 16 is a polyethylene glycol group
- the solvent of the compound S1 solution is dimethyl sulfoxide.
- the solvent is hydrazine, hydrazine-dimethylacetamide.
- the influence of the concentration of the probe molecule solution and the soaking time on the coverage of the probe molecule on the gold film was measured.
- the probe molecule ⁇ 2 was taken as an example, and the results are shown in the following table: 30 minutes 7% 15% 35% 45% 50%
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JP2004251807A (ja) * | 2003-02-21 | 2004-09-09 | Nipro Corp | エンドトキシン測定用センサ、測定方法、診断方法、製造方法およびセンサ再生方法 |
JP2006095452A (ja) * | 2004-09-30 | 2006-04-13 | Fuji Photo Film Co Ltd | スピンコート製膜法 |
CA2667394A1 (en) * | 2006-10-24 | 2008-06-12 | Rna Holding B.V. | Immobilisation and application of antigenic carbohydrates to detect infective micro-organisms |
CN101776637B (zh) * | 2009-01-13 | 2013-11-06 | 国家纳米科学中心 | 一种光电化学生物传感器及其制备方法 |
CN101887017B (zh) * | 2010-06-07 | 2012-01-11 | 北京理工大学 | 一种表面等离子体共振传感器芯片及其制备方法 |
WO2012009239A2 (en) * | 2010-07-13 | 2012-01-19 | The University Of Houston System | Sensors and separation based on molecular recognition via electropolymerization and colloidal layer templates |
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2012
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US20030107741A1 (en) * | 2001-12-11 | 2003-06-12 | Pyo Hyeon Bong | Surface plasmon resonance sensor system |
CN101591711A (zh) * | 2009-03-16 | 2009-12-02 | 唐国林 | 肽核酸探针生物芯片及其表面等离子体共振的检测方法 |
US8243276B2 (en) * | 2009-11-30 | 2012-08-14 | Corning Incorporated | Variable penetration depth biosensor |
CN102923968A (zh) * | 2012-11-13 | 2013-02-13 | 中国科学院理化技术研究所 | 一种表面等离子体共振传感芯片及其制备方法、应用 |
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CN111829991A (zh) * | 2020-08-04 | 2020-10-27 | 东北大学 | 基于β-CD的反射式光纤胆固醇传感器及其制备方法 |
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EP2921463A4 (en) | 2016-10-26 |
JP6329595B2 (ja) | 2018-05-23 |
CN102923968A (zh) | 2013-02-13 |
JP2016197122A (ja) | 2016-11-24 |
EP2921463A1 (en) | 2015-09-23 |
US20160266038A1 (en) | 2016-09-15 |
CN102923968B (zh) | 2015-06-10 |
JP2015535592A (ja) | 2015-12-14 |
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