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Sensor for detecting 8-hydroxyl-2'-deoxyguanosine as well as synthesis method and application thereof

A deoxyguanosine and sensor technology, applied in the fields of sensor synthesis and photochemical analysis, can solve problems such as limited scientific knowledge, and achieve huge market benefits, great application value, and simple preparation.

Inactive Publication Date: 2018-07-20
LIAOCHENG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Certain components of air pollution have been previously studied to induce or increase oxidative stress in biological systems, but scientific knowledge of the exact mechanisms is limited

Method used

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  • Sensor for detecting 8-hydroxyl-2'-deoxyguanosine as well as synthesis method and application thereof
  • Sensor for detecting 8-hydroxyl-2'-deoxyguanosine as well as synthesis method and application thereof
  • Sensor for detecting 8-hydroxyl-2'-deoxyguanosine as well as synthesis method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Synthetic method of 8-hydroxy-2'-deoxyguanosine sensor sensor

[0059] Two copies of 5 μL (microliter) of magnetic nanoparticles having a mass concentration of 1 mg / mL that have been modified with streptavidin were taken respectively, and a deoxynucleotide sequence chain ( probe1 and probe2) 7.5 μL (microliter), then dilute the two solutions into 1 mL of buffer solution, and react at 30°C for 20 minutes; after the reaction, mix the two solutions evenly, and then add 7.5 μL of a deoxynucleotide sequence (aptamer) solution with a molar concentration of 100nM / mL that can specifically bind to 8-hydroxy-2'-deoxyguanosine, and react at 30°C for 20 minutes, so that the 8-hydroxy - 2'-deoxyguanosine sensor.

[0060] figure 2 It can be seen that when the concentration of 8-hydroxy-2'-deoxyguanosine reaches pM (linear range 30pM–10.0nM), the reaction is extremely sensitive. Since the change value of the resonance light scattering signal is inversely proportional to the molecu...

Embodiment 2

[0208] Using the 8-hydroxy-2'-deoxyguanosine sensor prepared in Example 1. 3.0nM 8-hydroxy-2'-deoxyguanosine, the concentration is 100nM / L creatinine (creatinine), bovine serum albumin (BSA), ascorbic acid (ascorbic acid), sodium oxalate (disodium oxalate), sucrose (sucrose) , urea, uric acid, glucose, 2'-deoxyguanosine, ganciclovir, β-thymidine, guanine ), Mg 2+ 、K + , Ca 2+ , Fe 3+ 、Cu 2+ Test the resonant light scattering signal respectively.

[0209] Figure 8 Shown is a synthetic 8-hydroxy-2'-deoxyguanosine sensor that detects 8-hydroxy-2'-deoxyguanosine even when other biomolecules are present at higher concentrations than 8-hydroxy-2'-deoxyguanosine , still will not affect the detection of 8-hydroxy-2'-deoxyguanosine. So the 8-hydroxy-2'-deoxyguanosine sensor has good selectivity to 8-hydroxy-2'-deoxyguanosine.

Embodiment 3

[0211] Sensor reuse

[0212] After the 8-hydroxy-2'-deoxyguanosine sensor prepared in Example 1 was placed on the magnetic separator for 24 hours, the magnetic microspheres were tightly attracted by the magnetic separator. Carefully remove the supernatant, add a solution of a deoxynucleotide sequence (aptamer) that can specifically bind to 8-hydroxy-2'-deoxyguanosine, so that the concentration is 0.75nmol / mL, and finally dilute the whole system into 2mL of buffer solution, shake well and react at 30°C for 20 minutes, thus assembling a reusable 8-hydroxy-2'-deoxyguanosine sensor. By analogy, the same situation, the sensor can be reused.

[0213] Figure 9 The cycle is shown, and the high and low points on each broken line represent the resonance light scattering intensity before and after adding 8-hydroxy-2'-deoxyguanosine. It shows that the sensor performs well in repeated use.

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Abstract

The invention belongs to the technical field of sensor synthesis and the field of photochemical analysis, and in particular relates to a sensor which takes superparamagnetic nanoparticles capable of being circularly utilized as a carrier and aims at detecting 8-hydroxyl-2'-deoxyguanosine, a synthesis method of the sensor and application of the sensor to the fields of the photochemical analysis andenvironmental chemistry. The sensor is composed of superparamagnetic ferric oxide nanoparticles covered with streptavidin, and three different biotin modified single-chain deoxynucleotide sequences;one of the deoxynucleotide sequences is a deoxynucleotide sequence capable of specifically bind to the 8-hydroxyl-2'-deoxyguanosine; the streptavidin covers the superparamagnetic ferric oxide nanoparticles; the streptavidin is connected with the biotin, and the biotin is connected with the deoxynucleotide sequences. The sensor has good selectivity on the 8-hydroxyl-2'-deoxyguanosine, has strong anti-interference performance, and can be repeatedly utilized.

Description

technical field [0001] The invention belongs to the technical field of sensor synthesis and the field of photochemical analysis, in particular to a sensor for detecting 8-hydroxy-2'-deoxyguanosine (8-OHdG) with recyclable superparamagnetic nanoparticles as a carrier As well as its synthesis method and its application in the fields of photochemical analysis and environmental chemistry. Background technique [0002] 8-Hydroxy-2'-deoxyguanosine (8-OHdG) is a product of oxidative stress in which hydroxyl radicals attack the C8 site of guanosine in DNA sequences. This biomarker of oxidative stress occurs when the normal ratio of oxidants to antioxidants is out of balance. 8-Hydroxy-2'-deoxyguanosine is hypermutagenic and, although not directly lethal to affected cells, is a significant promoter of cancer. Furthermore, increased levels of 8-hydroxy-2'-deoxyguanosine have been linked to neurological disorders such as Alzheimer's disease, Parkinson's disease and atherosclerosis as...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N21/47
CPCG01N21/47G01N33/5308
Inventor 岳巧丽陶丽霞王永平李晨钟
Owner LIAOCHENG UNIV
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