Fluorescent nanometer molecular imprinting biomimetic sensor, preparation method and applications thereof

A technology of molecular imprinting and fluorescent nanometers, applied in fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of fast, economical, convenient, high-sensitivity and high-specificity detection of alpha-fetoprotein, achieve good molecular recognition ability, shorten Time, accuracy and precision results

Inactive Publication Date: 2018-01-19
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Technical problem to be solved: Aiming at the difficult problem of rapid, economical, convenient, high-sensitivity and high-specificity detection of the tum

Method used

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  • Fluorescent nanometer molecular imprinting biomimetic sensor, preparation method and applications thereof
  • Fluorescent nanometer molecular imprinting biomimetic sensor, preparation method and applications thereof
  • Fluorescent nanometer molecular imprinting biomimetic sensor, preparation method and applications thereof

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Experimental program
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Effect test

Embodiment 1

[0032] (1) Add 1.051 g of citric acid and 335 μL of ethylenediamine into 20 mL of ultrapure water, stir to dissolve; transfer the resulting solution to a 50 mL polytetrafluoroethylene autoclave, and react at 200 ° C for 5 h; naturally cool to room temperature, and pure Dialyzed in water for 48 hours, and freeze-dried for 12 hours to obtain carbon dots for future use.

[0033] (2) Ultrasonic dispersion of 20 mg of carbon dot powder in 20 mL of phosphate buffer solution with pH=5, followed by adding 100 μL of 1M EDC and NHS, and magnetic stirring for 20 minutes; adding 300 μL of 0.25M 4-VPA to the above solution for covalent condensation reaction, Magnetic stirring reaction 4h. After the reaction, the reaction solution was dialyzed in water for 48 hours to remove unreacted 4-VPA, other reactants and ions. Freeze-dry for 12 hours to obtain carbon dots modified with double bonds on the surface, which are ready for use.

[0034] (3) Add 1.25mg / mL AFP 160μL, 0.5mmol NIPAAm and 4-V...

Embodiment 2

[0037] (1) Add 1.051 g of citric acid and 335 μL of ethylenediamine into 20 mL of ultrapure water, stir to dissolve; transfer the resulting solution to a 50 mL polytetrafluoroethylene autoclave, and react at 200 ° C for 5 h; naturally cool to room temperature, and pure Water dialysis for 48 hours, and freeze-drying for 12 hours to obtain carbon dots for future use.

[0038] (2) Ultrasonic dispersion of 20 mg of carbon dot powder in 20 mL of phosphate buffer solution with pH=5, followed by adding 200 μL of 1M EDC and NHS, and magnetic stirring for 20 minutes; adding 300 μL of 0.25M 4-VPA to the above solution for covalent condensation reaction, Magnetic stirring reaction 4h. After the reaction was completed, the reaction solution was dialyzed in water for 48 hours to remove unreacted 4-VPA, other reactants and ions. Freeze-dry for 12 hours to obtain carbon dots modified with double bonds on the surface, which are ready for use.

[0039] (3) Add 1.25mg / mL AFP 320μL, 0.5mmol NI...

Embodiment 3

[0042] (1) Add 1.051 g of citric acid and 335 μL of ethylenediamine into 20 mL of ultrapure water, stir to dissolve; transfer the resulting solution to a 50 mL polytetrafluoroethylene autoclave, and react at 200 ° C for 5 h; naturally cool to room temperature, and pure Water dialysis for 48 hours, and freeze-drying for 12 hours to obtain carbon dots for future use.

[0043] (2) Ultrasonic dispersion of 20mg carbon dot powder in 20mL pH=5 phosphate buffer solution, then add 200μL 1M EDC and NHS in turn, and magnetic stirring reaction for 20min; add 300μL 0.25M 4-VPA to the above solution for covalent condensation reaction, magnetic The reaction was stirred for 4h. After the reaction was completed, the reaction solution was dialyzed in water for 48 hours to remove unreacted 4-VPA, other reactants and ions. Finally, freeze-dry for 12 hours to obtain carbon dots modified with double bonds on the surface, which are ready for use.

[0044] (3) Take 1.25mg / mL AFP 320μL, 0.5mmol NIP...

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Abstract

The invention relates to a fluorescent nanometer molecular imprinting biomimetic sensor, a preparation method and applications thereof. The preparation method comprises: weighing carbon quantum dot powder, ultrasonically dispersing in a borate buffer solution, sequentially adding 1-ethyl-(3-(dimethylamino)propyl)carbodiimide hydrochloride and N-hydroxysuccinimide, carrying out a stirring reactionat a room temperature under a dark condition, adding 4-vinylaniline, continuously carrying out the stirring reaction, carrying out dialysis on the obtained product in water, and carrying out freeze drying to obtain surface double bond functionalized carbon quantum dots; adding a template molecule and two different functional monomers to a pore forming agent, ultrasonically dissolving, and carryingout stirring pre-polymerization at a room temperature to obtain a pre-assembly solution A; ultrasonically dispersing the surface double bond functionalized carbon quantum dots in a pore forming agentto obtain a solution B; uniformly mixing the solution A and the solution B, adding a cross-linking agent and an initiator, introducing nitrogen, stirring, carrying out centrifugation, collecting theprecipitate, and washing with distilled water; and finally carrying out elution on the template protein by using HAc-SDS, and carrying out freeze drying on the obtained product so as to obtain the fluorescent nanometer molecular imprinting biomimetic sensor.

Description

technical field [0001] The invention belongs to the field of fluorescent nano-molecular imprint bionic sensors, in particular to a pH- and temperature-sensitive fluorescent nano-molecular imprint bionic sensor for liver cancer tumor markers and its preparation method and application. Background technique [0002] Liver cancer is a malignant tumor of the liver. It is one of the most common malignant tumors in the world. Its incidence rate ranks fifth and its mortality rate ranks third. The main reason for the extremely high mortality rate of liver cancer is that it is already at an advanced stage when it is diagnosed clinically. Therefore, early detection and early diagnosis of liver cancer are very important for the successful cure of cancer and the improvement of patient survival rate. At present, alpha-fetoprotein (alphafetoprotein, AFP), as a specific indicator of liver cancer, is an extremely important marker for the discovery of primary liver cancer. AFP is a glycoprot...

Claims

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Application Information

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IPC IPC(8): G01N21/64
Inventor 陈立娜孙成红黄姣姣姚丹丹顾小丽魏芳弟
Owner NANJING MEDICAL UNIV
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