Fluorescent molecularly imprinted polymer based on glycopeptide, preparation method and application in glycoprotein screening and detection

A fluorescent molecular imprinting and polymer technology, applied in the field of biosensing and bioanalysis, can solve the problems of complex structure, variability, large molecules, etc., and achieve the effect of improving specificity, binding force and good photostability

Active Publication Date: 2021-07-30
XI AN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to deficiencies such as flexible protein conformation, complex structure, large molecule, and variability in organic solvents, its application in MIPs is limited.

Method used

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  • Fluorescent molecularly imprinted polymer based on glycopeptide, preparation method and application in glycoprotein screening and detection
  • Fluorescent molecularly imprinted polymer based on glycopeptide, preparation method and application in glycoprotein screening and detection
  • Fluorescent molecularly imprinted polymer based on glycopeptide, preparation method and application in glycoprotein screening and detection

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preparation example Construction

[0042] Preparation of water-soluble N-GQDs. Dissolve CA (0.22-0.23 g) and EDA (0.18-0.22 mL) in ultrapure water (4.5-5.5 mL), and stir for 10-30 minutes until clear. The solution was transferred to a 20mL polytetrafluoroethylene reactor, and reacted at 160-200°C for 6-10h. Excess absolute ethanol was added to the obtained product to obtain solid N-GQDs.

[0043] In order to prepare N-GQDs@SiO 2 For nanoparticles, add N-GQDs (10-12mL), TEOS (20-60μL) and APTES (20-60μL) into absolute ethanol (30-50mL) under magnetic stirring, and react at room temperature for 1-3h. After washing three times with absolute ethanol, it was dried with a freeze dryer.

[0044] Subsequently, for the N-GQDs@SiO 2 Amino groups and double bonds are modified on the surface of nanoparticles, and N-GQDs@SiO 2 (790-810mg), anhydrous toluene (10-11mL), APTES (5-6mL) and MPS (5-6mL) were put into a round bottom flask, sonicated for 30min to obtain a suspension. the suspension in N 2 Reflux at 90-120°C ...

Embodiment 1

[0052] 1. Materials and methods

[0053] 1. Experimental materials

[0054] OVA, bovine serum albumin (BSA), bovine hemoglobin (BHb) and pronase E (Pronase E) (Beijing Suolaibao Technology Co., Ltd.); horseradish peroxidase (HRP) and trypsin (Tryspin) ( Shanghai Sangon Bioengineering); α-methacrylic acid (MAA), 3-aminopropyltriethoxysilane (APTES), 3-methacryloyloxypropyltrimethoxysilane (MPS), tetraethyl Oxysilane (TEOS), tetramethylethylenediamine (TEMED) and N, N'-methylenebisacrylamide (MBA) (Shanghai McLean Biochemical Technology Co., Ltd.); 4-formylphenylboronic acid (4- FPBA) (Shanghai Aladdin Biochemical Technology Co., Ltd.); ethylenediamine (EDA) and sodium borohydride (NaBH 4 ) (Shaanxi Sinopharm Holding Chemical Reagent Co., Ltd.); ammonium bicarbonate (NH 4 HCO 3 ), ammonium persulfate (APS), sodium chloride (NaCl) and sodium dodecyl sulfate (SDS) (Guangdong Guanghua Technology Co., Ltd.).

[0055] 2. Preparation of g-FMIPs

[0056] 2.1N-GQDs@SiO 2 Synthesi...

Embodiment 2

[0104] 2.1 N-GQDs@SiO 2 Synthesis of @FPBA nanoparticles

[0105] Preparation of water-soluble N-GQDs. CA (0.22 g) and EDA (0.18 mL) were dissolved in ultrapure water (4.5 mL), stirred for 10 minutes until clear. The solution was transferred to a 20mL polytetrafluoroethylene reactor, and reacted at 200°C for 6h. Excess absolute ethanol was added to the obtained product to obtain solid N-GQDs.

[0106]In order to prepare N-GQDs@SiO 2 For nanoparticles, N-GQDs (11 mL), TEOS (20 μL) and APTES (20 μL) were added into absolute ethanol (30 mL) under magnetic stirring, and reacted at room temperature for 3 h. After washing three times with absolute ethanol, it was dried with a freeze dryer.

[0107] Subsequently, for the N-GQDs@SiO 2 Amino groups and double bonds are modified on the surface of nanoparticles, and N-GQDs@SiO 2 (790mg), anhydrous toluene (10mL), APTES (5mL) and MPS (5mL) were put into a round bottom flask and ultrasonicated for 30min. the suspension in N 2 Refl...

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Abstract

The invention discloses a fluorescent molecularly imprinted polymer based on glycopeptide, a preparation method and application in glycoprotein screening and detection. The fluorescent molecularly imprinted polymer is prepared by the following steps of dispersing short glycopeptide immobilized N-GQDs@SiO2@FPBA of OVA in a phosphate buffer solution, adding MAA and MBA, performing ultrasonic dissolution, pre-assembling at 28-32 DEG C for 11-13 hours, then adding TEMED and APS, reacting at 28-32 DEG C for 5-7 hours under the protection of inert gas, eluting, and acquiring the fluorescent molecularly imprinted polymer based on the glycopeptide. Compared with a traditional glycoprotein detection method, the fluorescent molecularly imprinted polymer based on glycopeptide has the advantages of high specific recognition and sensitive and rapid detection, and screening and detection of trace target glycoprotein from a biological sample are very significant.

Description

technical field [0001] The invention belongs to the technical field of biosensing and bioanalysis, and relates to a glycopeptide-based fluorescent molecularly imprinted polymer, a preparation method and an application in glycoprotein screening and detection. Background technique [0002] Glycoproteins are very important parts of protein families involved in various cellular functions such as molecular recognition, signal transduction, cell adhesion and immune response. Clinical trials have found that human glycoprotein abnormalities are closely related to the occurrence of various cancers, such as alpha-fetoprotein and liver cancer, carcinoembryonic antigen and colon cancer or rectal cancer, etc., so they are often used as important biomarkers for disease diagnosis and targeted therapy things. However, the extremely low concentration of glycoproteins with important biological significance in biological samples and the interference of complex substances make their detection ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64C09K11/06C08F220/06C08F230/08C08F222/38C08J9/28C08L33/02
CPCG01N21/64G01N21/6428C09K11/06C08F220/06C08J9/28G01N2021/6417G01N2021/6432C09K2211/14C08J2333/02C08F230/085C08F222/385
Inventor 解笑瑜刘霞李静杨怡璇
Owner XI AN JIAOTONG UNIV
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