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Method for analyzing interaction between single carbon nanotube and protein

A protein interaction and carbon nanotube technology, applied in the field of biomolecular interaction, can solve the problems of unfavorable in situ observation, fluorescent label quenching and bleaching, and complicated sample preparation process, and achieve convenient and simple sample preparation and simple preparation method , The effect of simple detection method

Active Publication Date: 2019-05-14
UNIV OF SCI & TECH OF CHINA
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

However, scanning electron microscopy (SEM) and atomic force microscopy (AFM) can only image but cannot obtain binding kinetic curves, and ultimately cannot obtain relevant information such as association rate constants and dissociation rate constants.
However, total internal reflection fluorescence microscopy (TIRF) requires the use of fluorescent labeling, which often leads to complicated sample preparation processes, and fluorescent labeling will interfere with the status of normal samples, which is not conducive to in-situ observation.
In addition, fluorescent labels often suffer from quenching and bleaching, which prevents long-term observation

Method used

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  • Method for analyzing interaction between single carbon nanotube and protein
  • Method for analyzing interaction between single carbon nanotube and protein
  • Method for analyzing interaction between single carbon nanotube and protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: Interaction of bovine serum albumin and carbon nanotubes

[0056] Detect the interaction between 0.01, 0.1, 1.0mg / mL BSA and three MWNT-COOH

[0057] (1): first coat a 2-3nm thick chromium layer on the upper surface of the glass substrate, then coat a 40-50nm thick gold film on the upper surface of the chromium layer; place the gold film in 1mM SH-PEG- Soak in OH (M.W.=340) ethanol solution at 0°C for 24 hours to form a uniform PEG-OH (M.W.=340) SAM layer on the surface as an SPR sensor chip;

[0058] (2): Take 10 mg of carboxy-modified multi-walled carbon nanotubes (MWNT-COOH) and ultrasonically disperse them in ethanol solution for 5 min, pass the solution through a water phase filter membrane with d=0.22 μm, and take 10 μL of filtered supernatant for spin coating On the SPR sensor chip, the carbon nanotubes were confirmed to be single and uniformly dispersed by atomic force microscopy (AFM);

[0059] (3): Cover the small PDMS pool on the SPR sensor chip,...

Embodiment 2

[0077] Embodiment 2: Interaction of wheat germ agglutinin (WGA) and carbon nanotubes

[0078] Detection of the interaction between 0.1mg / mL WGA and three MWNT-COOH

[0079] (1): first coat a 2-3nm thick chromium layer on the upper surface of the glass substrate, then coat a 40-50nm thick gold film on the upper surface of the chromium layer; place the gold film in 1mM SH-PEG- Soak in OH (M.W.=340) ethanol solution at 0°C for 24 hours to form a uniform PEG-OH (M.W.=340) SAM layer on the surface as an SPR sensor chip;

[0080] (2): Take 10 mg of carboxy-modified multi-walled carbon nanotubes (MWNT-COOH) and ultrasonically disperse them in ethanol solution for 5 min, pass the solution through a water phase filter membrane with d=0.22 μm, and take 10 μL of filtered supernatant for spin coating On the SPR sensor chip, the carbon nanotubes were confirmed to be single and uniformly dispersed by atomic force microscopy (AFM);

[0081] (3): Cover the small PDMS pool on the SPR sensor ...

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Abstract

The invention belongs to the field of analyzing biomolecular interaction, and discloses a method for analyzing interaction between a single carbon nanotube and a protein. The method comprises the steps that after the carbon nanotube is pretreated, a plasma resonance sensing chip modified with a single molecular layer is spin-coated with the carbon nanotube, a protein solution is dispersed around the carbon nanotube, and a combined kinetic curve of the single carbon nanotube on the plasma resonance sensing chip and the protein solution is determined by utilizing intensity changes of surface plasmon resonance light. According to the method for analyzing the interaction between the single carbon nanotube and the protein, the tedious and complicated sample preparation by fluorescence labelingcan be eliminated on the basis of a surface plasmon resonance microscopy (SPRM) technology, and real-time label-free analysis imaging of in-situ interaction between different single carbon nanotubes and proteins is realized.

Description

technical field [0001] The invention belongs to the field of biomolecular interaction research, in particular to a method for analyzing the interaction between a single carbon nanotube and a protein, in particular to a method for analyzing a single carbon nanotube and a protein based on surface plasmon resonance imaging (SPRM) method of interaction. Background technique [0002] Carbon nanotubes, also known as bucky tubes, are one-dimensional nanomaterials with a special structure (the radial dimension is on the order of nanometers, the axial dimension is on the order of micrometers, and both ends of the tube are basically sealed). Carbon nanotubes are mainly coaxial tubes with several to tens of layers of carbon atoms arranged in a hexagonal shape. Carbon nanotubes can be regarded as curled graphene sheets, so according to the number of layers of graphene sheets, they can be divided into: single-walled carbon nanotubes and multi-walled carbon nanotubes. The interlayers ar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/552
Inventor 刘贤伟朱丽萍
Owner UNIV OF SCI & TECH OF CHINA
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