Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

An aptamer probe for detecting two tumor markers, an electrochemical biosensor, its preparation method and application

A tumor marker and aptamer technology, applied in the field of analytical chemistry, can solve problems affecting the analytical performance of sensors

Active Publication Date: 2020-07-03
GUANGZHOU YUEWANG AGRI CO LTD
View PDF9 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Whether the electrochemical aptamer sensor based on this principle is a signal-on type or a signal-off type, there is always a certain background current problem, which will affect the analytical performance of the sensor to a certain extent.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • An aptamer probe for detecting two tumor markers, an electrochemical biosensor, its preparation method and application
  • An aptamer probe for detecting two tumor markers, an electrochemical biosensor, its preparation method and application
  • An aptamer probe for detecting two tumor markers, an electrochemical biosensor, its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Optimization of assembly time of the aptamer probe of the present invention

[0056] In this embodiment, the method for optimizing the assembly time of the aptamer probe includes the following steps:

[0057] (1) Polish the gold electrode on the suede in 0.3 μm and 0.05 μm alumina slurry until it becomes a mirror surface, rinse with distilled water, and then ultrasonicate in secondary water, ethanol, and secondary water for 5 minutes, and then Soak the electrode in Piranha solution for 15min, rinse with secondary water and place the gold electrode in 0.1M H 2 SO 4 The cyclic voltammetry scan was performed between -0.3V and +1.5V, and then the cleaned gold electrode was rinsed with secondary water.

[0058] (2) 20 μL of 3.8 μM aptamer probe was dropped onto the surface of the pretreated gold electrode, and self-assembled at room temperature for different times (10 min, 20 min, 40 min, 80 min, 120 min, 160 min, overnight).

[0059] (3) After the gold electrod...

Embodiment 2

[0062] Embodiment 2 Hpa II endonuclease digestion time optimization

[0063] In the present embodiment, the Hpa II endonuclease digestion time optimization method comprises the following steps:

[0064] (1) Polish the gold electrode on the suede in 0.3 μm and 0.05 μm alumina slurry until it becomes a mirror surface, rinse with distilled water, and then ultrasonicate in secondary water, ethanol, and secondary water for 5 minutes, and then Soak the electrode in Piranha solution for 15min, rinse with secondary water and place the gold electrode in 0.1M H 2 SO 4 The cyclic voltammetry scan was performed between -0.3V and +1.5V, and then the cleaned gold electrode was rinsed with secondary water.

[0065] (2) 20 μL of 3.8 μM aptamer probe was dropped onto the surface of the pretreated gold electrode, and self-assembled at room temperature for 2 h.

[0066] (3) After the gold electrode was washed with secondary water, 20 μL of 1 mM mercaptohexanol was dropped onto the surface of ...

Embodiment 3

[0070] Embodiment 3Hpa II endonuclease concentration optimization

[0071] In the present embodiment, the Hpa II endonuclease concentration optimization method comprises the following steps:

[0072] (1) Polish the gold electrode on the suede in 0.3 μm and 0.05 μm alumina slurry until it becomes a mirror surface, rinse with distilled water, and then ultrasonicate in secondary water, ethanol, and secondary water for 5 minutes, and then Soak the gold electrode in Piranha solution for 15min, rinse with secondary water and put the gold electrode in 0.1M H 2 SO 4 The cyclic voltammetry scan was performed between -0.3V and +1.5V, and then the cleaned gold electrode was rinsed with secondary water.

[0073] (2) 20 μL of 3.8 μM aptamer probe was dropped onto the surface of the pretreated gold electrode, and self-assembled at room temperature for 2 h.

[0074] (3) After the gold electrode was washed with secondary water, 20 μL of 1 mM mercaptohexanol was dropped onto the surface of ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses an aptamer probe for detecting two kinds of tumor makers and an electrochemical biosensor and a preparation method and application thereof. The aptamer probe comprises an aptamer sequence, the aptamer sequence and the tumor marker I are subjected to specific recognition and closely bound, the 5' end and the 3' end of the aptamer probe are subjected to complementary hybridization to make the aptamer probe form a hairpin structure, and the stem portion of the hairpin structure comprises a recognition sequence of the tumor maker II and a recognition site of restriction enzyme. The electrochemical biosensor is prepared by fixing the aptamer probe on the surface of a working electrode. By means of the aptamer probe for detecting the two kinds of tumor makers and the electrochemical biosensor and the preparation method and application thereof, parallel analysis of multiple kinds of tumor marker molecules of thrombin and M.Sss I DNA methylation transferase can be achieved; the biosensor is high in detection sensibility, the lower detection limit for the thrombin is 0.1 ng / mL, and the lower detection limit for the M.Sss I DNA methylation transferase is 0.04 U / mL.

Description

technical field [0001] The invention relates to the field of analytical chemistry, in particular to an aptamer probe for detecting two tumor markers, an electrochemical biosensor and its preparation method and application, which can be used for the analysis and detection of major disease markers such as tumors. Background technique [0002] Clinically, rapid and sensitive screening of malignant tumors often uses tumor markers as detection indicators. Tumor markers are a class of substances produced by tumor tissues and cells that are related to tumor formation and occurrence, mainly including tumor antigens, hormones, enzymes and their isozymes. For example, thrombin is a serine proteolytic enzyme formed from the thrombin precursor. It can catalyze fibrinogen into fibrin, promote blood coagulation and regulate coagulation. Prognosis and other aspects are of great significance. In addition, DNA methyltransferase can catalyze DNA methylation, which plays an important role in...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/3277G01N27/48
Inventor 张松柏李玉红成诗琦杨基峰胡霞沈广宇卢基林
Owner GUANGZHOU YUEWANG AGRI CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com