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A method for the detection of glycoproteins by molecularly imprinted microspheres based on boronic acid fluorescent probes

A molecularly imprinted microsphere and fluorescent probe technology, applied in the field of glycoprotein detection, can solve the problem of time-consuming and other biomolecules such as fragile proteins, and achieve the effects of avoiding the sample processing process, mild reaction conditions and simple preparation method.

Inactive Publication Date: 2015-08-19
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, traditional mass spectrometry, chromatography and electrophoresis detection methods need to process biological samples before detection, and this process is extremely easy to destroy biomolecules such as proteins and takes too long. Therefore, it is necessary to develop a non-destructive, high-sensitivity and rapid detection method

Method used

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  • A method for the detection of glycoproteins by molecularly imprinted microspheres based on boronic acid fluorescent probes
  • A method for the detection of glycoproteins by molecularly imprinted microspheres based on boronic acid fluorescent probes
  • A method for the detection of glycoproteins by molecularly imprinted microspheres based on boronic acid fluorescent probes

Examples

Experimental program
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Effect test

Embodiment 1

[0029] 1. Amino groups bonded to the surface of monodisperse porous cross-linked glycidyl methacrylate microspheres

[0030] Add 2.0 g of monodisperse porous cross-linked glycidyl methacrylate microspheres into 50 mL of 1 mol / mL sulfuric acid aqueous solution, stir in a water bath at 60 ° C for 24 hours, centrifuge, remove the supernatant, and wash the product with deionized water to medium properties, vacuum drying at 60°C to obtain monodisperse cross-linked glycidyl methacrylate microspheres with hydroxyl groups on the surface; disperse 0.5g of the microspheres in 100mL of absolute ethanol, add 1mL of ammonia water and 20mL of 3-aminopropyl Triethoxysilane, ultrasonically dispersed for 0.5 hours, stirred at room temperature for 12 hours, centrifuged to remove the supernatant, the precipitate was washed with absolute ethanol, and dried in vacuum at 60°C to obtain a monodisperse cross-linked methacrylic epoxy with amino groups on the surface Propyl ester microspheres.

[0031...

Embodiment 2

[0043] Monodisperse crosslinked glycidyl methacrylate microspheres with boric acid fluorescent probes bonded to the surface were prepared according to steps 1-2 of Example 1. In step 3 of Example 1, the horseradish peroxidase used was replaced with an equal mass of ovalbumin to obtain ovalbumin molecularly imprinted microspheres with fluorescent properties. According to the method of embodiment 1 step 4, the fluorescence spectrogram of fluorescence intensity changing with ovalbumin concentration and the standard curve of fluorescence intensity changing with the logarithmic value of ovalbumin concentration at the maximum emission wavelength of 430nm are drawn, the results are shown in Figure 3-4 .

[0044] Depend on Figure 3-4 It can be seen that under the same detection conditions, the ovalbumin molecularly imprinted microspheres with fluorescent properties have obvious fluorescent responses to the template molecule ovalbumin. When the concentration of ovalbumin is 0.05-1.0...

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Abstract

The invention discloses a method for detecting glycoprotein by molecularly imprinted microspheres based on a boric acid fluorescence probe. The method comprises the following steps: with monodisperse poly(glycidylmethacrylate-co-ethylenedimethacrylate) microspheres as a carrier, grafting the boric acid fluorescence probe on the surface of the carrier; with the boric acid fluorescence probe as a functional monomer, and appointed glycoprotein as a template molecule, polymerizing the template molecule with the functional monomer to form a stable prepolymer; and preparing the appointed glycoprotein molecularly imprinted microspheres by adopting a surface imprint technology. The molecularly imprinted microspheres have selective recognition performance on the appointed glycoprotein, are capable of directly detecting the glycoprotein by adopting a fluorescence method, and are relatively simple to operate, and relatively high in detection speed in comparison with traditional mass-spectrography or chromatography detection.

Description

technical field [0001] The invention belongs to the technical field of glycoprotein detection, and in particular relates to a method for detecting glycoproteins by molecularly imprinted polymer microspheres based on boric acid fluorescent probes. Background technique [0002] Glycoproteins are an important class of post-translational modification proteins, which play important roles in molecular recognition, intracellular and intercellular signal transduction, and immune response. Almost all key molecules involved in innate and adaptive immunity in the human immune system are glycosylated proteins. Abnormal protein glycosylation is the culprit in the occurrence, development and metastasis of some tumor diseases. When tumor diseases occur, glycoproteins on the cell surface are overexpressed. Some glycoproteins, such as alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA), have been used as clinical markers to detect cancer. Therefore, the study of glycoproteins has im...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 张静倪艳丽魏静茹
Owner SHAANXI NORMAL UNIV
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