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Cathode photoelectrochemical detection model of T4 polynucleotide kinase and application of cathode photoelectrochemical detection model

A polynucleotide and photoelectrochemical technology, which is applied in the field of detection models for cathodic photoelectrochemical detection of T4 polynucleotide kinases, can solve the problems of interference, low biological reaction recognition efficiency, and complicated operation.

Active Publication Date: 2022-03-18
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Anode photoelectrochemical analysis is not only susceptible to the interference of reducing substances in the medium, but also the reported methods need to immobilize biomolecules on the electrode surface, which has high cost (biomolecules need to be labeled), cumbersome / time-consuming operation, and possible The photodamage of biomolecules and the low efficiency of biological reaction recognition

Method used

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  • Cathode photoelectrochemical detection model of T4 polynucleotide kinase and application of cathode photoelectrochemical detection model
  • Cathode photoelectrochemical detection model of T4 polynucleotide kinase and application of cathode photoelectrochemical detection model
  • Cathode photoelectrochemical detection model of T4 polynucleotide kinase and application of cathode photoelectrochemical detection model

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preparation example Construction

[0045] (1) Bi 2 o 3 Preparation of nanomaterials: 0.2-0.5g soluble bismuth salt (Bi(NO 3 ) 3 ·5H 2 O or bismuth sulfate) was dissolved in 60 mL of deionized water, then 0.5-0.9 g of trisodium citrate, 0.24 g of urea and 0.50-1.0 g of polyvinylpyrrolidone were added and vigorously stirred for 20-50 minutes; subsequently, the resulting mixed solution was transferred to Put it in a high-pressure reactor and react at 100-180°C for 5-12 hours; after cooling to room temperature, wash the obtained sample with ethanol and deionized water several times, and finally dry the sample at 60°C overnight to obtain basic carbonic acid Bismuth powder; finally, the obtained bismuth subcarbonate was heated in a muffle furnace at 350-450°C for 2 hours to obtain Bi 2 o 3 Powder samples of nanomaterials;

[0046] (2) Bi 2 o 3 / ITO electrode preparation: the Bi 2 o 3 The nanomaterials were dispersed in deionized water to make a suspension; then the resulting suspension was drop-coated on th...

Embodiment 1

[0051] A cathodic photoelectrochemical detection model of T4 polynucleotide kinase, its construction method comprises the steps:

[0052] a. Bi 2 o 3 Preparation of nanomaterials: Weigh 0.48g bismuth nitrate and dissolve it in 60mL deionized water, then add 0.88g trisodium citrate, 0.24g urea and 0.50g polyvinylpyrrolidone and stir vigorously for 30 minutes; subsequently, transfer the obtained mixed solution Put it in a high-pressure reactor, and react at 180° C. for 12 hours. After cooling to room temperature, the obtained sample was washed several times with ethanol and deionized water, and the final sample was dried overnight at 60°C to obtain bismuth subcarbonate powder; finally, the obtained bismuth subcarbonate was heated in a muffle furnace at 400°C Heated for 2 hours to obtain Bi 2 o3 Powder samples of nanomaterials; e.g. figure 1 shown above Bi 2 o 3 SEM images of nanomaterials; such as figure 2 shown above Bi 2 o 3 X-ray diffraction patterns of nanomaterial...

Embodiment 2

[0060] A cathodic photoelectrochemical detection model of T4 polynucleotide kinase, its construction method comprises the steps:

[0061] a. Bi 2 o 3 Preparation: 0.35 g of bismuth sulfate was dissolved in 60 mL of deionized water, then 0.88 g of trisodium citrate, 0.24 g of urea, and 1.0 g of polyvinylpyrrolidone were added and vigorously stirred for 30 minutes; subsequently, the resulting mixed solution was transferred to a high-pressure reaction In the kettle, react at 160°C for 10 hours; after cooling to room temperature, the obtained sample is washed several times with ethanol and deionized water, and the final sample is dried overnight at 60°C to obtain bismuth subcarbonate powder; finally, the obtained The bismuth subcarbonate was heated in a muffle furnace at 400°C for 2 hours to obtain Bi 2 o 3 Powder samples of nanomaterials;

[0062] b. Bi 2 o 3 / ITO electrode preparation: the prepared Bi 2 o 3 Nanomaterials were added to deionized water, and a 1.0 mg / mL sus...

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Abstract

The invention relates to the field of analysis and detection, in particular to a detection model for non-labeled and non-fixed cathode photoelectrochemical detection of T4 polynucleotide kinase as well as a preparation method and application of the detection model. The construction method of the detection model comprises the following steps: (1) preparation of a Bi2O3 nano material; (2) preparation of a Bi2O3 / ITO electrode; (3) preparation of biological reaction solutions with different T4 polynucleotide kinase concentrations; (4) determination of light current; when the detection model is used for detecting the T4 PNK, biomolecule immobilization is not needed, the operation is simple and convenient, the cost is low, the selectivity is good, the sensitivity is high, the linear range is wide (5.0 * 10 <-4 >-10 U / mL), the detection limit is low (1.0 * 10 <-4 > U / mL), and the detection model has huge potential in practical application.

Description

technical field [0001] The invention relates to the field of analysis and detection, in particular to a non-marked and non-fixed cathode photoelectrochemical detection model for detecting T4 polynucleotide kinase, a preparation method and application thereof. Background technique [0002] T4 polynucleotide kinase (T4 PNK) is an important kinase that can catalyze the phosphorylation of the 5' hydroxyl terminal, and is closely related to normal cell activities such as DNA recombination, replication, and damage repair[Li,P. -P.; Cao, Y.; Mao, C.-J.; Jin, B.-K.; Zhu, J.-J. Anal.Chem.2019, 91, 1563-1570.]. Therefore, the determination of the activity of T4 PNK is of great significance for clinical diagnosis, drug development and revealing of basic biological processes [Hou, T.; Wang, X.-Z.; Liu, X.-J.; Lu, T.-T.; Liu, S.-F.; Li, F. Anal. Chem. 2014, 86, 884-890.]. [0003] The conventional methods for determining T4 PNK mainly include isotope labeling [Bernstein, N.K.; Williams...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N1/28
CPCG01N27/3278G01N1/28
Inventor 王光丽赵玲玲董玉明陈彦如刘田利
Owner JIANGNAN UNIV
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