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Photoelectrochemical biosensor for detecting 5-hydroxymethylcytosine (5hmC) deoxyribonucleotide and preparation method thereof

A technology of deoxyribonucleotides and hydroxymethylcytosine, which is applied in the field of photoelectrochemical biosensors and its preparation, can solve the problems of expensive, cumbersome operation steps, and the need for professional operators, and achieve easy operation and high detection selectivity , The effect of improving the detection sensitivity

Inactive Publication Date: 2018-11-30
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods all require expensive instruments and cumbersome operation steps, and need professional operators.
Moreover, the detection of 5hmC by the above method is easily interfered by 5mC
There is no report on the detection of 5hmC by photoelectrochemical analysis method based on tungsten sulfide

Method used

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  • Photoelectrochemical biosensor for detecting 5-hydroxymethylcytosine (5hmC) deoxyribonucleotide and preparation method thereof
  • Photoelectrochemical biosensor for detecting 5-hydroxymethylcytosine (5hmC) deoxyribonucleotide and preparation method thereof
  • Photoelectrochemical biosensor for detecting 5-hydroxymethylcytosine (5hmC) deoxyribonucleotide and preparation method thereof

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preparation example Construction

[0050] (1) Preparation of polydopamine: Weigh 0.5-1 g of dopamine and dissolve it in 300-600 mL of deionized water. Under magnetic stirring, heat to 45-80°C. Then, 2-4 mL of NaOH solution with a concentration of 0.5-2 M was added. Stand for reaction for 3-8 hours. Centrifuge at 2000-5000rpm to collect the suspension. Then the suspension was centrifuged at 10000-15000 rpm to collect the precipitate. After the precipitate was washed 3-5 times with deionized water, it was vacuum-dried at 60°C.

[0051] (2) Preparation of tungsten sulfide dispersion: weigh 5-250 mg of tungsten sulfide, add it into 50 mL of deionized water, and ultrasonically disperse for 1-3 hours.

[0052] (3) Preparation of polydopamine dispersion: weigh 10-100 mg of polydopamine, add it into 20 mL of deionized water, and disperse by ultrasonic for 1-3 hours.

[0053] (4) ITO electrode pretreatment: divide the ITO conductive glass into 5×1cm 2 , then ultrasonically cleaned with ethanol / NaOH mixture (ratio ...

Embodiment 1

[0068] Embodiment 1: the preparation of polydopamine

[0069] Weigh 0.54 g of dopamine and dissolve it in 300 mL of deionized water. Under magnetic stirring, heat to 50°C. Then, 2 mL of 1 M NaOH solution was added. Stand for reaction for 5 hours. Centrifuge at 4000 rpm to collect the suspension. Then the suspension was centrifuged at 12000 rpm to collect the precipitate. The precipitate was washed three times with deionized water, and then dried under vacuum at 60°C.

Embodiment 2

[0070] Embodiment 2: the preparation of tungsten sulfide dispersion liquid

[0071] Weigh 12 mg of tungsten sulfide, add it into 3 mL of deionized water, and ultrasonically disperse for 1-3 hours.

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Abstract

The invention discloses a photoelectrochemical biosensor for detecting 5-hydroxymethylcytosine (5hmC) deoxyribonucleotide and a preparation method thereof. The photoelectrochemical biosensor comprisesan electrode and tungsten sulfide, polydopamine, mercaptobenzoic acid, 5hmC, phos-tag-biotin and streptavidin which are sequentially modified on the surface of the electrode. High sensitivity and high specificity detection of 5 hmC can be realized by use of the good photoelectric activity and biocompatibility of the tungsten sulfide, the excellent electronic donor properties and biocompatibilityof the polydopamine, the specific recognition and binding properties of phos-tag-biotin to phosphoric acid groups, and the specific reaction of hydroxymethyl and sulfydryl groups on the 5hmC catalyzedby M.HhaI methyltransferase. The method can well eliminate the interference of 5mC to the detection of the 5hmC. The detection method is simple, instrument miniaturization is realized, the photoelectrochemical biosensor is easy to operate, and the detection of the 5hmC can be realized by simple treating of the surface of the electrode.

Description

technical field [0001] The invention relates to the technical field of photoelectrochemical analysis, in particular to a photoelectrochemical biosensor for detecting 5-hydroxymethylcytosine deoxyribonucleotides and a preparation method thereof. Background technique [0002] 5-Hydroxymethylcytosine deoxyribonucleotide (5hmC) was first discovered in phage in 1952. It can be modified by glycosyltransferase-mediated glycosylation, so that the phage genome can resist the host after entering the host Degradation by restriction enzymes. But at the time this discovery did not attract enough attention. In 2009, researchers found that 5hmC was abundantly expressed in human, mouse brain and embryonic stem cells, and the concept of hydroxymethylation came into people's field of vision again and gained attention. Currently 5hmC is the "sixth base" after 5mC is called the "fifth base". Although the function of 5hmC is not yet fully understood, it may also be an important epigenetic mar...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/416G01N27/30
CPCG01N27/305G01N27/3275G01N27/4166
Inventor 殷焕顺周云雷艾仕云隋程吉王月王明慧
Owner SHANDONG AGRICULTURAL UNIVERSITY
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