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Fluorescent Velocity Sensing Method for Quantitative Detection of Glycosylated Proteins

A glycosylated protein and speed sensing technology, applied in the field of biomedicine, can solve the problems of large error value, poor detection stability and poor versatility, and achieve the effect of fast detection speed

Active Publication Date: 2018-01-30
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Tests have proved that although the above method can quickly determine the concentration of sugar / glycoprotein, the error value in the detection process is too large, and the detection stability and versatility are not good.

Method used

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  • Fluorescent Velocity Sensing Method for Quantitative Detection of Glycosylated Proteins
  • Fluorescent Velocity Sensing Method for Quantitative Detection of Glycosylated Proteins
  • Fluorescent Velocity Sensing Method for Quantitative Detection of Glycosylated Proteins

Examples

Experimental program
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Embodiment 1

[0034] In this embodiment, taking the detection of immunoglobulin G (hereinafter referred to as IgG) as an example, the FSS-CSE test is first performed on the prepared IgG standard samples of various concentrations to obtain the fluorescence sensing speeds corresponding to the standard samples of various concentrations, so that A standard curve of fluorescence sensing speed and protein concentration was constructed ( image 3 -B). After constructing the standard curve, the same method can be used to measure the IgG of unknown concentration. After measuring the fluorescence sensing speed of IgG of unknown concentration, the concentration value can be calculated according to the fluorescence sensing speed and the standard curve.

[0035] This embodiment specifically includes the following steps:

[0036] In the first step, take the quantitative determination of ovalbumin concentration as an example. In this embodiment, 5 concentration points are selected, which are 0.2, 0.5, 1...

Embodiment 2

[0047] In this embodiment, the ovalbumin in Example 1 is replaced by glycosylated hemoglobin (HbA1c) related to diabetes detection, and a standard curve can be obtained similarly, such as image 3 C shows. The content of glycosylated hemoglobin in human blood can be obtained using the same method as above.

[0048] The gel is also a polyacrylamide gel, and its concentration, cross-linking degree and components are consistent with those in the ovalbumin FSS-CSE test.

[0049] Both the background electrolyte solution and the electrode buffer are 20 mM pH 7.4 phosphate buffer solution.

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Abstract

A fluorescent speed sensing method for quantitatively detecting glycosylated protein is provided. FSS-CSE testing is performed on glycosylated protein standard products, corresponding different migration rates of fluorescein released after glycosylated protein in different concentrations is combined with fluorescent sensing boric acid probe F1@Rp3 in an electric field are obtained, and a linear relation between the glycosylated protein in known concentration and the migration rates of fluorescein is established; then the glycosylated protein in unknown concentration is tested through the same FSS-CSE, the corresponding migration rates of the fluorescein in the electric field are acquired, and the corresponding migration rates of the fluorescein are substituted in the linear relation so that the concentration of the glycosylated protein can be acquired. The glycated hemoglobin (HbA1c) in blood can be fast and quantitatively measured. Besides, the detection speed is high, and only half an hour is needed for measuring the concentration of the glycosylated protein.

Description

technical field [0001] The invention relates to a technique in the field of biomedicine, in particular to a fluorescence velocity sensing method for quantitatively detecting glycosylated proteins. Background technique [0002] Protein glycosylation is an important post-translational modification, which is closely related to many life activities of organisms. Glycosylated proteins represent most cell surface markers and secreted proteins, and about 50% to 60% of proteins in the human body are modified by glycosylation. The research on glycoprotein (glycoprotein, GP) is beneficial to the identification of biological markers and therapeutic targets of diseases, and the discovery of potential therapeutic targets, thus providing assistance for disease diagnosis. [0003] Glycosylated protein analysis methods include chromatography, affinity chromatography, zone electrophoresis, capillary electrophoresis, ultraviolet spectrophotometry, electrochemical methods, high performance li...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 曹成喜吴雪静孟庆华樊柳荫肖华丛峰松
Owner SHANGHAI JIAOTONG UNIV
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