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Fluorescence method for detecting beta-lactoglobulin based on quantum dot-nucleic acid aptamer-graphene oxide

A nucleic acid aptamer, lactoglobulin technology, applied in the field of biological nanotechnology sensing, can solve the problems of transient evanescence and easy quenching, and achieve the effects of strong stability, improved sensitivity, and strong photobleaching threshold.

Pending Publication Date: 2020-07-17
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional fluorescent dyes show shortcomings such as easy quenching and fleeting in the detection process, and quantum dots have unique advantages

Method used

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  • Fluorescence method for detecting beta-lactoglobulin based on quantum dot-nucleic acid aptamer-graphene oxide
  • Fluorescence method for detecting beta-lactoglobulin based on quantum dot-nucleic acid aptamer-graphene oxide
  • Fluorescence method for detecting beta-lactoglobulin based on quantum dot-nucleic acid aptamer-graphene oxide

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Embodiment 1-9

[0038] A fluorescent method for detecting β-lactoglobulin based on quantum dot-nucleic acid aptamer-graphene oxide, comprising the following steps: a modified carboxyl group activation method on the water-soluble quantum dot, the steps are as follows:

[0039] The carboxyl-modified quantum dots were dissolved in 1 mmol / L MES buffer with EDC and NHS at a ratio of 1:1:1.5, and activated by shaking at room temperature for 20 min.

[0040] The activated carboxyl quantum dots and amino-modified nucleic acid aptamer link method, the steps are as follows:

[0041] The amino-modified aptamer was dissolved in PBS buffer (pH 7.4), aptamer: quantum dot = 10:1, and reacted by shaking on a shaker at room temperature for 48 hours. After the quantum dots were labeled, the mixed system was purified by ultrafiltration twice.

[0042] The method for binding the target object to the quantum dot-aptamer fluorescent probe, the steps are as follows:

[0043] Add different concentrations of β-lact...

Embodiment 10

[0051] A fluorescent method for detecting β-lactoglobulin based on quantum dot-nucleic acid aptamer-graphene oxide, comprising the steps of:

[0052] A modified carboxyl group activation method on water-soluble quantum dots, the steps are as follows:

[0053] The carboxyl-modified quantum dots were dissolved in 1 mmol / L MES buffer with EDC and NHS at a ratio of 1:1:1.5, and activated by shaking at room temperature for 20 min.

[0054] The activated carboxyl quantum dots and amino-modified nucleic acid aptamer link method, the steps are as follows:

[0055] The amino-modified aptamer was dissolved in PBS buffer (pH7.4), aptamer:quantum dot=12:1, and reacted on a shaker at room temperature for 48 hours. After the quantum dots were labeled, the mixed system was purified by ultrafiltration three times.

[0056] The method for binding the target object to the quantum dot-aptamer fluorescent probe, the steps are as follows:

[0057] Add different concentrations of β-lactoglobulin...

Embodiment 11

[0060] A fluorescent method for detecting β-lactoglobulin based on quantum dot-nucleic acid aptamer-graphene oxide, comprising the steps of:

[0061] A modified carboxyl group activation method on water-soluble quantum dots, the steps are as follows:

[0062] The carboxyl-modified quantum dots were dissolved in 1 mmol / L MES buffer with EDC and NHS at a ratio of 1:1:1.5, and activated by shaking at room temperature for 20 min.

[0063] The activated carboxyl quantum dots and amino-modified nucleic acid aptamer link method, the steps are as follows:

[0064] The amino-modified aptamer was dissolved in PBS buffer (pH 7.4), aptamer:quantum dot=5:1, and reacted on a shaker at room temperature for 48 hours. After the quantum dots were labeled, the mixed system was purified by ultrafiltration three times.

[0065] The method for binding the target object to the quantum dot-aptamer fluorescent probe, the steps are as follows:

[0066]Add different concentrations of β-lactoglobulin ...

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Abstract

The invention relates to a fluorescence method for detecting beta-lactoglobulin based on quantum dot-nucleic acid aptamer-graphene oxide. Carboxyl modified water-soluble quantum dots are used as a fluorophore; an amino-modified aptamer is used as a recognition molecule; graphene oxide (GO) is used as a quenching agent, a novel composite nano fluorescent probe is constructed, a reactant is shot andimaged under an ultraviolet lamp, and quantitative detection data is obtained by using image J image processing software, thereby realizing high-sensitivity detection of target molecule beta-lactoglobulin.

Description

technical field [0001] The invention belongs to the field of biological nanotechnology sensing, and in particular relates to a fluorescence method for detecting β-lactoglobulin based on quantum dot-nucleic acid aptamer-graphene oxide. Background technique [0002] β-lactoglobulin (β-Lg) is a milk-specific protein synthesized by mammary epithelial cells and is the main whey protein component in ruminant and pig milk. Studies in recent years have shown that human milk also contains β-lactoglobulin, but the content is very small, and gastric acid and pepsin cannot digest and hydrolyze it, so it can enter the blood circulation through the gastrointestinal tract. β-lactoglobulin accounts for 43.6%-50.0% of the total protein in bovine milk whey, is the most abundant protein in whey, and the content in skimmed milk is about 2.0-4.0g / L. The β-lactoglobulin in milk has a molecular weight of about 18400Da and an isoelectric point of 5.1-5.3. It consists of 162 amino residues and has ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/68G01N33/533
CPCG01N21/64G01N21/6428G01N21/6456G01N33/68G01N33/533G01N2021/6417G01N2021/6432G01N2021/6441G01N2333/4713
Inventor 陆旸张咏王芮王硕
Owner TIANJIN UNIV OF SCI & TECH
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