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Fluorescence biosensor, and preparation method and purpose thereof

A biosensor and fluorescence technology, applied in the field of biosensors, can solve the problems of limiting detection sensitivity and detection range, ignoring and reducing background signals, etc., and achieve the effects of low cost, good selectivity and good biocompatibility

Active Publication Date: 2017-03-08
ANHUI NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most of them use signal cycle amplification to improve sensitivity, ignoring the reduction of background signal, which limits the detection sensitivity and detection range

Method used

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  • Fluorescence biosensor, and preparation method and purpose thereof
  • Fluorescence biosensor, and preparation method and purpose thereof
  • Fluorescence biosensor, and preparation method and purpose thereof

Examples

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Embodiment 1

[0083] A method based on two-dimensional manganese dioxide nanosheets--a fluorescence sensing platform constructed by a fluorescent probe, comprising the following steps:

[0084] (1) Prepare 10mM potassium permanganate solution and 0.1M morpholineethanesulfonic acid buffer solution with pH=6.0 for synthesizing manganese dioxide nanosheets.

[0085] (2) Dissolve the purchased FAM-DNA sequence (FAM-DNA: FAM-AAA AAA AAA AAA AAA) in 0.1M Tris-HCl (pH 7.4) buffer solution, and store it at 4°C for later use; Phosphatase was dissolved in 0.01M Tris-HCl buffer solution containing 5mM magnesium chloride and 0.1mM zinc chloride, pH=8.12, and stored at -4°C for use.

[0086] (3), 5ml potassium permanganate solution was added to 12ml morpholine ethanesulfonic acid buffer solution, the mixed solution was ultrasonicated for 30 minutes to form a tan solution, then the ultrasonically mixed solution was centrifuged at 7000rpm for 15 minutes, and deionized water Wash the precipitate with etha...

Embodiment 2

[0096] A method based on two-dimensional manganese dioxide nanosheets--a fluorescence sensing platform constructed by a fluorescent probe, comprising the following steps:

[0097] Add 40ul FAM-DNA solution to the manganese dioxide nanosheet solution containing 0.2mg / ml, mix well, and place it at 37°C for 5min to attenuate the fluorescence of FAM-DNA, then add vitamin C magnesium phosphate and alkaline phosphatase, React in a thermostat at 37°C for 30 minutes to fully react magnesium vitamin C phosphate and alkaline phosphatase to generate ascorbic acid, reduce manganese dioxide nanosheets, release FAM-DNA, restore fluorescence, and obtain an "on" fluorescent biosensor .

[0098] A method and application of a fluorescent sensing platform based on two-dimensional manganese dioxide nanosheets-fluorescent probes, the specific method is:

[0099] a. Add 40ul FAM-DNA solutions to a series of manganese dioxide nanosheet solutions containing 0.2mg / ml, mix well, and place at 37°C for ...

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Abstract

The invention relates to a fluorescence biosensor, and a preparation method and a purpose thereof. On the basis of a two-dimensional manganese dioxide nanometer sheet as a fluorescence sensing platform, morpholineethanesulfonic acid is used for reducing potassium permanganate for synthesizing the manganese dioxide nanometer sheets; the manganese dioxide nanometer sheets adsorb FAM-ssDNA and quench FAM fluorescence; under the effect of alkaline phosphatase, vitamin C magnesium L-ascorbate-2-phosphate is hydrolyzed to generate ascorbic acid; the two-dimensional manganese dioxide nanometer sheets are reduced into manganese ions by the ascorbic acid; FAM-ssDNA is released; the fluorescence is recovered; the fluorescence sensor using the manganese dioxide nanometer sheets and the alkaline phosphatase to regulate the fluorescence intensity is built and is used for detecting the activity of the alkaline phosphatase. Compared with the prior art, the fluorescence biosensor has the characteristics of simplicity, high speed, low background signal, high sensitivity, low detection limit and good selectivity by using the change of the two-dimensional manganese dioxide nanometer sheets on the fluorescence signal, i.e., the off-on mode for detecting the alkaline phosphatase.

Description

technical field [0001] The invention belongs to the technical field of biosensors, and in particular relates to a method and application of a fluorescent sensing platform constructed based on two-dimensional manganese dioxide nanosheets-fluorescent probes, which can realize sensitive detection of alkaline phosphatase activity. Background technique [0002] Alkaline phosphatase (ALP) is a phosphomonoesterase that promotes the dephosphorylation of various phosphorylated substrates. Alkaline phosphatase has been shown to play a vital role in the regulation of intracellular processes such as cell growth, apoptosis, and is therefore often used as a diagnostic indicator for various diseases, including breast and prostate cancer, bone disease, abnormal liver function ,diabetes. In addition, alkaline phosphatase also plays an important role in environmental and food quality monitoring. Therefore, it is extremely important to design a simple and sensitive method for detecting alkal...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6432
Inventor 王广凤朱升美
Owner ANHUI NORMAL UNIV
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