A kind of method of screening Helicobacter pylori urease inhibitor

A technology for urease inhibitors and Helicobacter pylori, which is applied in the field of screening Helicobacter pylori urease inhibitors to achieve the effects of simple operation, easy storage, and easy operation

Active Publication Date: 2022-04-12
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Research and professional methods for screening Helicobacter pylori urease inhibitors based on biosensor technology are still blank

Method used

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  • A kind of method of screening Helicobacter pylori urease inhibitor
  • A kind of method of screening Helicobacter pylori urease inhibitor
  • A kind of method of screening Helicobacter pylori urease inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1: A kind of method of screening Helicobacter pylori urease inhibitor AHA

[0045] (1) Preparation and characterization of HPUb / nPt / NPG / GCE biosensor:

[0046] The NPG film prepared by corroding the Au / Ag alloy sheet with concentrated nitric acid at 40°C for 1 h was attached to the pretreated GCE; then nPt was electrodeposited on the surface of NPG / GCE by cathodic reduction plating to obtain nPt / NPG / GCE, in which The electrolyte contains 5mM H 2 PtCl 6 0.5mol / L H 2 SO 4 solution with a current density of 5mA / cm 2 , the electrodeposition time was 120s; finally, 20 μL of purified HPUb was drop-coated on the surface of the nPt / NPG / GCE, and self-assembled at 4°C for 24 hours; the HPUb / nPt / NPG / GCE biosensor was finally obtained;

[0047] The prepared HPUb / nPt / NPG / GCE was used as the working electrode, the platinum electrode was used as the counter electrode, and the saturated calomel electrode was used as the reference electrode to form a three-electrode syst...

Embodiment 2

[0060] Embodiment 2: A kind of method of screening Helicobacter pylori urease inhibitor hydroxyurea

[0061] (1) Preparation and characterization of HPUb / nPt / NPG / GCE biosensor:

[0062] The NPG film prepared by corroding the Au / Ag alloy sheet with concentrated nitric acid at 40°C for 1 h was attached to the pretreated GCE; then nPt was electrodeposited on the surface of NPG / GCE by cathodic reduction plating to obtain nPt / NPG / GCE, in which The electrolyte contains 5mM H 2 PtCl 6 0.5mol / L H 2 SO 4 solution with a current density of 5mA / cm 2 , the electrodeposition time was 120s; finally, 20 μL of purified HPUb was drop-coated on the surface of the nPt / NPG / GCE, and self-assembled at 4°C for 24 hours; the HPUb / nPt / NPG / GCE biosensor was finally obtained;

[0063] The prepared HPUb / nPt / NPG / GCE was used as the working electrode, the platinum electrode was used as the counter electrode, and the saturated calomel electrode was used as the reference electrode to form a three-electr...

Embodiment 3

[0072] Embodiment 3: A kind of method of screening Helicobacter pylori urease inhibitor methylurea

[0073] (1) Preparation and characterization of HPUb / nPt / NPG / GCE biosensor:

[0074] The NPG film prepared by corroding the Au / Ag alloy sheet with concentrated nitric acid at 40°C for 1 h was attached to the pretreated GCE; then nPt was electrodeposited on the surface of NPG / GCE by cathodic reduction plating to obtain nPt / NPG / GCE, in which The electrolyte contains 5mM H 2 PtCl 6 0.5mol / L H 2 SO 4 solution with a current density of 5mA / cm 2 , the electrodeposition time was 120s; finally, 20 μL of purified HPUb was drop-coated on the surface of the nPt / NPG / GCE, and self-assembled at 4°C for 24 hours; the HPUb / nPt / NPG / GCE biosensor was finally obtained;

[0075] The prepared HPUb / nPt / NPG / GCE was used as the working electrode, the platinum electrode was used as the counter electrode, and the saturated calomel electrode was used as the reference electrode to form a three-electro...

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Abstract

The invention discloses a method for screening Helicobacter pylori urease inhibitors using a HPUb / nPt / NPG / GCE biosensor. The steps include 1) preparation and characterization of the HPUb / nPt / NPG / GCE biosensor; / NPG / GCE biosensor detects the response current generated by urea with gradient concentration of 0-100mM, and calculates the K of urea detected by the biosensor according to the Mie equation a，尿素 and V max，尿素 3) Detecting different urease inhibitors, screening and analyzing the inhibition type and inhibition constant of each inhibitor. The HPUb / nPt / NPG / GCE biosensor developed in the method of the present invention has a simple preparation process, high detection stability, and is easy to store. At the same time, compared with other detection methods, the screening method provided by the present invention does not require complicated sample pretreatment processes. Moreover, the operation is simple, the detection is rapid, the screening time of potential urease inhibitors is effectively shortened, and the application prospect is broad.

Description

technical field [0001] The present invention relates to a kind of method of screening Helicobacter pylori (Helicobacter pylori, H. pylori) urease inhibitor, relate in particular to a method utilizing H. pylori urease b subunit (H. pylori 26695 urease b subunit, HPUb) / nano platinum (Nano Platinum, nPt) / nanoporous gold (Nanoporous gold, NPG) / glassy carbon electrode (Glassy carbon electrode, GCE) biosensor screening method for Helicobacter pylori urease inhibitors. It belongs to the field of electrochemical analysis and testing. Background technique [0002] Helicobacter pylori (H. pylori) is a Gram-negative microaerophilic bacterium that selectively colonizes the surface of human gastric mucosal epithelial cells and the bottom layer of gastric mucus. It is well known that H. pylori is associated with the development of chronic gastritis, peptic ulcer, gastric mucosa-associated lymphoid tissue lymphoma and gastric adenocarcinoma. Although combination therapies based on an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327C12Q1/34C12Q1/00
CPCG01N27/3277C12Q1/34C12Q1/005G01N2333/98
Inventor 王霞肖洒
Owner SHANDONG UNIV
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