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Guanosine tetraphosphate electrochemical nano-enzyme sensor and preparation method thereof

A guanosine tetraphosphate, electrochemical technology, applied in scientific instruments, instruments, material analysis through electromagnetic means, etc., can solve the problems of complex sensor preparation, long analysis time, instability, etc., and achieve high detection sensitivity and easy operation Simple and stable effect

Pending Publication Date: 2022-04-12
TIANJIN UNIV OF COMMERCE
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Liquid chromatography-mass spectrometry has high detection sensitivity and good accuracy, but it needs to rely on expensive instruments and professional testers, and the analysis time is relatively long
Chemical fluorescent probe technology has extremely high detection sensitivity, but the preparation of the sensor is complicated and unstable

Method used

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  • Guanosine tetraphosphate electrochemical nano-enzyme sensor and preparation method thereof
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  • Guanosine tetraphosphate electrochemical nano-enzyme sensor and preparation method thereof

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Embodiment 1

[0036] 1. Pretreatment of glassy carbon electrodes:

[0037] Polish the glassy carbon electrode in an 8-shape on the suede with alumina slurry with a particle size of 0.03 μm, wash it with water, and then wash it in an ultrasonic water bath for tens of seconds until the surface is smooth and clean; the electrode is cleaned at 1×10 -3 mol / L K 3 Fe(CN) 6 Solution (containing 0.20mol / LKNO 3 ), the electrode is activated by cyclic voltammetry with a scanning range of 0.6-0.1V and a scanning speed of 50mV / s, and the scanning is repeated until a stable cyclic voltammetry curve appears. The above-mentioned stable cyclic voltammetry curve meets the following requirements: the peak potential difference should be below 80mV, and as close as possible to 64mV, the electrode can be used, and finally placed in a nitrogen environment to dry for use. Compared with the conventional glassy carbon electrode pretreatment method, the operation of the present invention is simpler and the require...

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Abstract

The invention discloses a guanosine tetraphosphate electrochemical nano-enzyme sensor and a preparation method thereof, and aims to provide a sensor capable of biologically detecting ppGpp and a preparation method thereof. According to the sensor, gold nanoparticles and guanosine tetraphosphate hydrolase MESH1 molecules are adsorbed on a substrate through a bridging agent chitosan, and the substrate is sealed through a sealing liquid, so that a chitosan film layer, a gold nanoparticle sol layer, an MESH1 enzyme layer and a sealing layer are formed on the substrate. The preparation method comprises the following steps: adding a low-concentration chitosan solution on a pretreated electrode, and after drying, forming a chitosan film on the surface of the electrode; airing the electrode with the chitosan membrane, placing the electrode in the nanogold sol, and carrying out self-assembly at 4 DEG C for at least 12 hours; washing with ultrapure water, airing, and self-assembling in an MESH1 zymoprotein solution at 4 DEG C for at least 12 hours; sealing non-specific sites by using sealing liquid after cleaning; and cleaning and airing to obtain the sensor. The sensor is high in sensitivity, strong in specificity, rapid in quantification and short in response time.

Description

technical field [0001] The invention relates to the technical field of biological detection, more specifically, relates to a guanosine tetraphosphate electrochemical nano-enzyme sensor and a preparation method thereof, the application of the guanosine tetraphosphate electrochemical nano-enzyme sensor, and the use of the obtained A method for the biological detection of guanosine tetraphosphate by an electrochemical nano-enzyme sensor for guanosine tetraphosphate. Background technique [0002] Guanosine tetraphosphate (ppGpp) is an important compound that transmits nitrogen starvation signals in organisms, and plays a vital role in the growth and survival of organisms. Currently known ppGpp detection methods include: based on liquid chromatography-mass spectrometry (UPLC-MS) and chemical fluorescent probe technology. Liquid chromatography-mass spectrometry has high detection sensitivity and good accuracy, but it needs to rely on expensive instruments and professional testers...

Claims

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Application Information

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IPC IPC(8): G01N27/416G01N27/327
Inventor 鲁丁强庞广昌任瑞娟裴陈琳刘丹阳冯春雷王轼
Owner TIANJIN UNIV OF COMMERCE
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