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Molecular biosensor and method of single-molecule detection of DNA or protein using the same

A technology of biosensors and devices, applied in the direction of electrochemical variables of materials, etc., can solve the problems of difficult real-time detection, high sensitivity, selectivity, and lack of

Inactive Publication Date: 2012-03-14
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, many existing methods are difficult to perform real-time detection or kinetic analysis, some methods face high technical requirements, and some lack high sensitivity and selectivity

Method used

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  • Molecular biosensor and method of single-molecule detection of DNA or protein using the same
  • Molecular biosensor and method of single-molecule detection of DNA or protein using the same
  • Molecular biosensor and method of single-molecule detection of DNA or protein using the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: Application of single-molecule aptamer device to detect Thrombin

[0050] 1. Preparation of single-molecule aptamer devices

[0051] 1) On a highly conductive silicon chip substrate containing a thermal oxide layer of silicon dioxide with a thickness of 300nm (resistivity 5-20ohm cm -1 ), use chemical vapor deposition to grow high-quality ultra-long carbon nanotube arrays, and sequentially vapor-deposit Cr (5nm) and Au (50nm) on a single-walled carbon nanotube (7000 μm in length) at an interval of about 20 microns. As the source and drain of the device, construct the SWNT transistor device;

[0052] 2) The SWNT device obtained in step 1) is subjected to ultra-fine electron beam etching and oxidation cutting, and a nano-gap of 1-10 nm is obtained between the carbon tubes, figure 1 Figure b in the middle shows the SEM and AFM pictures of the device. The diameter of the carbon tube in the figure is ~1.2nm, and the size of the nanogap is ~10nm;

[0053] 3) In...

Embodiment 2

[0073] Example 2, the application of single-molecule aptamer device to detect immunoglobulin E (IgE)

[0074] Immunoglobulin E is a class of allotropic antibodies with a δ chain, and is the main antibody involved in the regulation of the pathogenesis of allergic rhinitis, allergic asthma, and eczema. The detection of IgE is of great significance to the study of allergic diseases.

[0075] 1. Preparation of single-molecule aptamer devices

[0076] 1) On a highly conductive silicon chip substrate containing a thermal oxide layer of silicon dioxide with a thickness of 300nm, use chemical vapor deposition to grow high-quality ultra-long carbon nanotube arrays, in a single-walled carbon nanotube (length Cr (5nm) and Au (50nm) were successively vapor-deposited at intervals of about 20 microns (7000 microns) as the source and drain of the device to construct a SWNT transistor device;

[0077] 2) The SWNT device obtained in step 1) is subjected to ultra-fine electron beam etching an...

Embodiment 3

[0087] Example 3, Application of single-molecule aptamer device to detect vascular endothelial growth factor (VEGF)

[0088] Vascular endothelial growth factor is a specific heparin-binding growth factor for vascular endothelial cells, which can induce angiogenesis in vivo and directly affect the content of VEGF in the process of tumor growth and spread. The detection of VEGF can be applied to the diagnosis and treatment of cancer.

[0089] 1. Preparation of single-molecule aptamer devices

[0090] 1) On a highly conductive silicon chip substrate containing a thermal oxide layer of silicon dioxide with a thickness of 300nm, use chemical vapor deposition to grow an array of high-quality ultra-long carbon nanotubes, in a single-walled carbon nanotube ( Cr (5nm) and Au (50nm) were successively vapor-deposited at intervals of about 20 microns on the length of 7000 microns as the source and drain of the device to construct a SWNT transistor device;

[0091] 2) The SWNT device obta...

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Abstract

The invention discloses a rapid and high sensitive molecular biosensor and a method of single-molecule detection of DNA or protein using the molecular biosensor. The molecular biosensor disclosed herein comprises: a) at least a single-walled carbon nanotube transistor device, wherein, the single-walled carbon nanotube transistor device comprises a grid electrode, a source electrode, a drain electrode and a conducting channel, the conducting channel is a single-walled carbon nanotube which is cut into two sections to form two single-walled carbon nanotubes with a gap of 1-10 nm; and b) an aptamer, wherein, the terminals of the aptamer are modified by terminal amino groups, and the aptamer is connected with the two ends of the carbon nanotubes forming the gap through an amide bond. The method is a reliable and practical method with general applicability. According to the method, nano / molecular electronics and the biosystem are combined, and the direct and real-time detection of the activities of DNA or protein with single-minded selectivity and ultrahigh sensitivity is realized by using functionalized mono-molecular devices and electric signal measurement.

Description

technical field [0001] The invention relates to a fast and highly sensitive molecular biosensor and a method for direct and real-time single-molecule detection of DNA or protein. Background technique [0002] Establishing a practical platform for direct detection of biological and chemical activities at the single-molecule level is one of the common goals of industry and scientific research, because it is important for a series of applications such as environmental monitoring, industrial quality control, and clinical diagnosis. significance. In order to reliably detect DNA-protein interactions, a variety of direct and labeled techniques have been developed such as: ELISA, surface plasmon resonance, electrochemistry, scanning probe microscopy, nanoparticles, microcantilever , carbon nanotubes, impedance, nanowires, etc. However, many existing methods are difficult to perform real-time detection or kinetic analysis. Some methods face high technical requirements, while others...

Claims

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Application Information

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IPC IPC(8): G01N27/26
Inventor 郭雪峰刘松
Owner PEKING UNIV
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