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Electrochemical biosensor for cell epithelial-mesenchymal transition detection as well as preparation method and application of electrochemical biosensor

A biosensor, electrochemical technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve the problems of cumbersome steps, high cost, long detection time, etc., and achieve the effect of high sensitivity

Active Publication Date: 2020-10-30
SHANDONG NORMAL UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, the inventors have found that the above method has defects including long detection time, cumbersome steps, and high cost.

Method used

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  • Electrochemical biosensor for cell epithelial-mesenchymal transition detection as well as preparation method and application of electrochemical biosensor
  • Electrochemical biosensor for cell epithelial-mesenchymal transition detection as well as preparation method and application of electrochemical biosensor
  • Electrochemical biosensor for cell epithelial-mesenchymal transition detection as well as preparation method and application of electrochemical biosensor

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preparation example Construction

[0040] In yet another specific embodiment of the present invention, the preparation method of the E-cad-antibody-QD probe comprises: adding E-cad-antibody and coupling agent 1-(3-dimethyl Aminopropyl)-3-ethylcarbodiimide hydrochloride was subjected to conjugation reaction and obtained after purification.

[0041] In yet another specific embodiment of the present invention, the specific method of purification is to pass the conjugate through a 0.2 μm filter membrane to remove large aggregates, and wash at least 5 times with borate buffer.

[0042] In yet another specific embodiment of the present invention, a method for preparing the above-mentioned electrochemical biosensor for detection of cell epithelial-mesenchymal transition is provided, the preparation method comprising:

[0043] Preparation of carboxylated carbon nanotube-gold nanoparticles modified electrode; and, preparation of E-cad-antibody-QD probe.

[0044] In yet another specific embodiment of the present inventi...

Embodiment 1

[0062] 1. Chemicals and reagents

[0063] Carboxylated carbon nanotubes (CNTs, diameter 30-50 nm, average length 10-20 μm) were purchased from Xfnano MaterialsTech (Nanjing, China). Qdot™ 655ITK™ carboxyl quantum dots (Q21321MP) were obtained from Thermo Fisher Scientific (Waltham, MA, USA). Boric acid, potassium chloride (KCl) and phosphate were purchased from Sinopharm Group Chemical Reagent Co.LTD (China). Anti-E-cad antibody, anti-N-cad antibody, anti-ZO-1 antibody, anti-vimentin antibody were provided by antibody cam. E-cadherin protein and chloroauric acid (HAuCl 4 ) were purchased from Sigma-Aldrich. Prepare all assay systems using double distilled water.

[0064] 2. Cell culture and EMT model construction

[0065] A549, PANC1 and MCF7 cells were cultured in DMEM medium supplemented with 10% FBS. RBE and SKOV3 cells were cultured in RPMI1640 medium supplemented with 10% FBS. All cells were purchased from the Cell Bank of the Chinese Academy of Sciences. Cells we...

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Abstract

The invention provides an electrochemical biosensor for cell epithelial-mesenchymal transition detection as well as a preparation method and application of the electrochemical biosensor, which belongto the technical field of molecular biology and biosensor preparation. The electrochemical biosensor prepared by the invention can specially detect the change of the marker protein (E-cadherin) of theEMT, and can effectively analyze different stages of the EMT. Due to the fact that molecular information is transmitted to the electronic equipment, signals of the EMT are greatly amplified. Meanwhile, due to the synergistic effect of the QD and carbon nano tube-gold nano particles, when the differential pulse voltammetry is used for detection, the response of electrochemical signals is rapid andsensitive, the electrochemical biosensing technology can be effectively promoted to be applied to research of complex biological behaviors, and therefore a good practical application value is achieved.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and biosensor preparation, and in particular relates to an electrochemical biosensor for detection of cell epithelial-mesenchymal transition and its preparation method and application. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Epithelial-mesenchymal transition (EMT) plays an important role in various biological processes, such as embryonic development, tissue growth, and wound healing, among others. More and more evidences show that EMT plays a key role in tumor progression, and EMT can promote the infiltration of benign tumor cells into surrounding tissues and metastasis to dis...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N27/26G01N27/30G01N27/327G01N33/533
CPCG01N21/6486G01N27/26G01N27/30G01N27/3278G01N33/533
Inventor 周军杜欣郑晓迪张振华
Owner SHANDONG NORMAL UNIV
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