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Nano-particle enhancement detection device based on non-modified monolayer graphene being used as working electrode and application thereof

A single-layer graphene and nanoparticle technology, applied in the field of detection, can solve problems such as low conduction electrons, and achieve the effects of improving detection efficiency, saving resources and improving accuracy

Active Publication Date: 2013-12-04
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] When using nanomaterials to detect biomolecules, including carbon nanotubes, graphene oxide and other materials as detection electrodes, it is necessary to functionalize the surface of these nanomaterials, such as carboxyl and other functional groups on the surface, so that they can bind to biomolecules , but there are many defects on the surface of materials such as modified graphene oxide, which causes the speed of conducting electrons to be far lower than that of single-layer graphene. The present invention relates to single-layer graphene that does not need to be modified as a detection electrode

Method used

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  • Nano-particle enhancement detection device based on non-modified monolayer graphene being used as working electrode and application thereof
  • Nano-particle enhancement detection device based on non-modified monolayer graphene being used as working electrode and application thereof
  • Nano-particle enhancement detection device based on non-modified monolayer graphene being used as working electrode and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0108] (1) Preparation of gold nanoparticles (diameter 20nm) coated with the detected protein and HRP:

[0109] First, prepare colloidal gold according to the method of Yu et al. (Yu et al., 2012), synthesize gold nanoparticles with a diameter of 20nm, and then take 200ul prepared nano-gold colloid solution (diameter 20nm, 6.8×10 ‐9 mol / L), add 2ul of K 2 CO 3 (wt.0.1%) to adjust its pH=8.9, then add 1ul (1mg / ml) CEA antibody and 1ul (1mg / ml) HRP, let it stand at room temperature for half an hour, centrifuge (9000rmp, 50min), remove the supernatant, Resuspend with PBS (pH7.4), add BSA (0.1%), and store at 2‐8°C. The ultraviolet spectrum of the obtained nano-gold is as figure 1 shown.

[0110] (2) Preparation of magnetic beads coated with captured protein:

[0111] Take 50ul, 10mg / ml carboxylated magnetic beads (Invitrogen, Cat.no.650.11 / 12), wash twice with an equal volume of MES (25mM, pH6.0), each time for 10 minutes, magnetically separate, and remove the supernatant ;...

Embodiment 2

[0115] (1) Preparation of gold nanoparticles (diameter 19nm) coated with the protein to be tested and HRP: First, gold nanoparticles with a diameter of 20nm were synthesized according to the method of colloidal gold prepared by Yu et al. (Yu et al., 2012), and then Take 200ul prepared nano-gold colloid solution (diameter 20nm, 6.8×10 ‐10 mol / L), add 2ul of K 2 CO 3 (wt.0.1%) to adjust its pH=9.2, then add 0.9ul (1.1mg / ml) of CEA antigen and 0.9ul (1.1mg / ml) of HRP, let stand at room temperature for half an hour, centrifuge (9000rmp, 50min), Remove the supernatant, resuspend with PBS (pH7.0), add BSA (0.5%), and store at 2‐8°C.

[0116] (2) Preparation of magnetic beads coated with captured protein:

[0117] Take 50ul, 10mg / ml carboxylated magnetic beads (Invitrogen, Cat.no.650.11 / 12), wash twice with an equal volume of MES (25mM, pH6.0), each time for 10 minutes, magnetically separate, and remove the supernatant Add 60ul of freshly prepared EDC (40mg / ml) and 60ul of NHS (4...

Embodiment 3

[0121] (1) Preparation of gold nanoparticles (diameter 21nm) coated with the detected protein and HRP:

[0122] First, prepare colloidal gold according to the method of Yu et al. (Yu et al., 2012), synthesize gold nanoparticles with a diameter of 20nm, and then take 200ul prepared nano-gold colloid solution (diameter 20nm, 6.8×10 ‐8 mol / L), add 2ul of K 2 CO 3 (wt.0.1%) to adjust its pH=8.6, then add 1.1ul (0.9mg / ml) mouse anti-human IgG antibody and 1.1ul (0.9mg / ml) HRP, let stand at room temperature for half an hour, centrifuge (9000rmp, 50min), remove the supernatant, resuspend with PBS (pH7.8), add BSA (0.1%), and store at 2‐8°C.

[0123] (2) Preparation of magnetic beads coated with captured protein:

[0124]Take 50ul, 10mg / ml carboxylated magnetic beads (Invitrogen, Cat.no.650.11 / 12), wash twice with an equal volume of MES (25mM, pH6.0), each time for 10 minutes, magnetically separate, and remove the supernatant ;Add freshly prepared 40ul EDC (60mg / ml) and 40ul NHS (...

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Abstract

The invention relates to the field of detection, in particular to a nano-particle enhancement detection device based on non-modified monolayer graphene being used as a working electrode and the application thereof, provides the application of the non-modified monolayer graphene in the field of nano-particle enhancement detection, and further provides the nano-particle enhancement detection device, a nano-particle enhancement detection method and a relevant kit. According to the nano-particle enhancement detection method, the non-modified graphene is used as the working electrode, and therefore the electronic transmission rate is improved; modified nanogold and magnetic beads are used for amplifying current signals, and detection sensitivity is improved.

Description

technical field [0001] The invention relates to the field of detection, in particular to a nanoparticle-enhanced detection device based on unmodified single-layer graphene as a working electrode and its application. Background technique [0002] The appearance of graphene has stirred up huge waves in the scientific community. People have discovered that graphene has extraordinary electrical conductivity, strength dozens of times stronger than steel, and excellent light transmission. Its appearance is expected to be widely used in modern electronic technology. The field sparked a revolution. In graphene, electrons can move extremely efficiently, while traditional semiconductors and conductors, such as silicon and copper, do not perform as well as graphene. Due to the collision of electrons and atoms, traditional semiconductors and conductors release some energy in the form of heat. At present, general computer chips waste 72%-81% of electric energy in this way. Graphene is d...

Claims

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Application Information

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IPC IPC(8): G01N27/30G01N27/48
Inventor 毛红菊金兵张宏莲金庆辉赵建龙
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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