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Nano immunological biosensor

A biosensor and nanotechnology, applied in the direction of biological testing, instruments, scientific instruments, etc., can solve the problems of non-reusable use, achieve high sensitivity, and simple preparation methods

Inactive Publication Date: 2004-08-25
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ben-Dov etc. (Ben-Dov, I., Willner, I., Zisman, E.Anal.Chem.1997,69,3506) adopt maleimide monolayer membrane to immobilize antibody, but this sensor are disposable and cannot be reused

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The immobilization of a gold nanocrystal on the surface of a gold electrode:

[0019] 1) Immerse the cleaned gold electrode in 0.1M 4-mercaptoaniline solution for 1 hour at room temperature, take it out and clean it with absolute ethanol to clean the 4-mercaptoaniline physically adsorbed on the surface of the electrode. Finally, rinse with a large amount of high-purity water and blow dry with high-purity nitrogen.

[0020] 2) Put the electrode into a gold nanocrystal solution with a pH value of 3, assemble it in the dark for 1 hour, and clean it with high-purity water after taking it out.

[0021] Immobilization of secondary antibodies on gold nanocrystal modified electrodes:

[0022] 1) Put the electrode modified with gold nanocrystals into 0.5ml phosphate buffer solution containing 0.1mg / ml antibody, pH is 6. And after mixing thoroughly for 20 minutes in a micro-vortex mixer, soak overnight at 4°C.

[0023] 2) After taking it out, use 0.02M phosphate buffer solutio...

Embodiment 2

[0027] The immobilization of a gold nanocrystal on the surface of a gold electrode:

[0028] 1) Immerse the cleaned gold electrode in 0.1M 4-mercaptoaniline solution for 4 hours at room temperature, take it out and clean it with absolute ethanol to clean the 4-mercaptoaniline physically adsorbed on the surface of the electrode. Finally, rinse with a large amount of high-purity water and blow dry with high-purity nitrogen.

[0029] 2) Put the electrode into a gold nanocrystal solution with a pH value of 10, assemble it in the dark for 5 hours, and clean it with high-purity water after taking it out.

[0030] Immobilization of secondary antibodies on gold nanocrystal modified electrodes:

[0031] 1) Put the electrode modified with gold nanocrystals into 0.5ml phosphate buffer solution containing 0.1mg / ml antibody, pH is 6. And after fully mixing for 20 minutes in a micro-vortex mixer, soak overnight at 37°C.

[0032] 2) After taking it out, use 0.02M phosphate buffer solution...

Embodiment 3

[0036] The immobilization of a gold nanocrystal on the surface of a gold electrode:

[0037] 1) Immerse the cleaned gold electrode in 0.1M 4-mercaptoaniline solution for 6 hours at room temperature, take it out and clean it with absolute ethanol to clean the 4-mercaptoaniline physically adsorbed on the electrode surface. Finally, rinse with a large amount of high-purity water and blow dry with high-purity nitrogen.

[0038] 2) Put the electrode into a gold nanocrystal solution with a pH value of 8, assemble it in the dark for 10 hours, and clean it with high-purity water after taking it out.

[0039] Immobilization of secondary antibodies on gold nanocrystal modified electrodes:

[0040] 1) The electrode modified with gold nanocrystals is placed in 0.5 ml of phosphate buffer solution containing 0.1 mg / ml antibody, and the pH is 7.4. And after fully mixing for 20 minutes in a micro-vortex mixer, soak overnight at 37°C.

[0041] 2) After taking it out, use 0.02M phosphate buf...

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Abstract

The invention chemically decorates the gold electrode surface with mercapto compound, like mercapto aniline, mercapto ethylamine, etc. It uses the mercapto end of the mercapto compound to combine with the gold electrode, chemically, and fixes the mercapto end on the gold electrode to make the amido on the other end exposed on the electrode surface. On a certain condition, the exposed amido can combine with the nano gold crystal by chemical bond, thus fixing the nano gold crystal on the gold electrode surface. The surface of the nano gold crystal has charges and can strongly absorb the antibody on the neutral or physiological pH condition, making the antibody fixed on the surface of the nano gold crystal, and thus making the nano immune biosensor.

Description

technical field [0001] The invention relates to a preparation method of a nanometer immune biosensor. Background technique [0002] Using chemical methods to detect the interaction between antigen and antibody is the development direction of many researches. Biosensors that combine high-sensitivity sensing technology with specific immune responses to monitor antigen and antibody responses are called immunosensors, and are used in medical diagnosis and treatment, environmental pollution monitoring, and food analysis. The working principle of the immunosensor is similar to that of the traditional immunoassay, which is a solid-phase immunoassay, that is, the antigen or antibody is immobilized on the surface of a solid support to detect the antibody or antigen in the sample. During the past 15 years of development in the field of immunodiagnostics, more interest has been placed on the binding kinetics of antibodies immobilized on the surface of solid supports. Biological sampl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/02G01N27/30G01N27/327G01N33/53
Inventor 李景虹王美佳孙春燕
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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