Preparation method and application of immunosensor based on ferritin three-dimensional structure

An immune sensor and three-dimensional structure technology, applied in the field of biosensing and new nanomaterials, can solve the problems of poor reproducibility and low sensitivity, and achieve low cost, high sensitivity and good reproducibility

Inactive Publication Date: 2019-09-13
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, electrochemical biosensors are mainly used to detect procalcitonin, but this method has the disadvantages of poor reproducibility and low sensitivity.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1. A preparation method and application of an immunosensor based on the three-dimensional structure of ferritin, comprising the following steps:

[0025] (1) Polish the glassy carbon electrode with a diameter of 4 mm with 1.0 μm, 0.3 μm, and 0.05 μm alumina polishing powder in sequence, and rinse it with ultrapure water;

[0026] (2) Drop-coat 6 μL of antibody-labeled isoluminol-ferritin-nanogold solution at a concentration of 2 mg / mL on the surface of the glassy carbon electrode as the sensor substrate material, and place it to dry at 37 °C;

[0027] (3) Add 3 μL bovine serum albumin solution with a mass fraction of 1% dropwise to block the non-specific active sites on the electrode surface, rinse the electrode surface with pH 7.4 phosphate buffer solution PBS, and place it at 4 °C C to dry;

[0028] (4) Add 6 μL standard solution of procalcitonin or procalcitonin solution of unknown concentration, incubate at 37 °C for 0.5 h, rinse the electrode surface wi...

Embodiment 2

[0029] Embodiment 2. A preparation method and application of an immunosensor based on the three-dimensional structure of ferritin, comprising the following steps:

[0030] (1) Polish the glassy carbon electrode with a diameter of 4 mm with 1.0 μm, 0.3 μm, and 0.05 μm alumina polishing powder in sequence, and rinse it with ultrapure water;

[0031] (2) Drop-coat 6 μL of antibody-labeled isoluminol-ferritin-nanogold solution at a concentration of 3 mg / mL on the surface of the glassy carbon electrode as the sensor substrate material, and place it to dry at 37 °C;

[0032] (3) Add 3 μL bovine serum albumin solution with a mass fraction of 2% dropwise to block the non-specific active sites on the electrode surface, rinse the electrode surface with pH 7.4 phosphate buffer solution PBS, and place it at 4 °C C to dry;

[0033] (4) Add 6 μL standard solution of procalcitonin or procalcitonin solution of unknown concentration, incubate at 37 °C for 1 h, wash the electrode surface with ...

Embodiment 3

[0034] Embodiment 3. A preparation method and application of an immunosensor based on the three-dimensional structure of ferritin, comprising the following steps:

[0035] (1) Polish the glassy carbon electrode with a diameter of 4 mm with 1.0 μm, 0.3 μm, and 0.05 μm alumina polishing powder in sequence, and rinse it with ultrapure water;

[0036] (2) Drop-coat 6 μL of antibody-labeled isoluminol-ferritin-nanogold solution at a concentration of 4 mg / mL on the surface of the glassy carbon electrode as the sensor substrate material, and place it to dry at 37 °C;

[0037] (3) Add 3 μL bovine serum albumin solution with a mass fraction of 3% dropwise to block the non-specific active sites on the electrode surface, rinse the electrode surface with pH 7.4 phosphate buffer solution PBS, and place it at 4 °C C to dry;

[0038] (4) Add 6 μL standard solution of procalcitonin or procalcitonin solution of unknown concentration, incubate at 37 °C for 2 h, wash the electrode surface with ...

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Abstract

The invention relates to a preparation method and application of an immunosensor based on a ferritin three-dimensional structure, belonging to the field of novel nano materials and the technical fieldof biosensing. The electrochemiluminescence (ECL) technology is adopted, ferritin is taken as a substrate, ECL luminescent molecules N-(4-Aminobutyl)-N-ethylisoluminol molecules are subjected to covalent cross-linking at the outer portion of the ferritin to obtain isoluminol-ferritin, a reaction condition is regulated and controlled, and the nanogold is reduced at the surface of the isoluminol-ferritin in situ to obtain a novel three-dimensional hybrid nano structure isoluminol-ferritin-nanogold. The present invention further provides a preparation method of an immunosensor based on the structure. The method is simple in preparation, low in cost, low in reaction energy consumption and environmental friendly, the sensor is used for detection of procalcitonin, the sensitivity is high, the specificity is high, the reproducibility is good, the detection limit is 3.4 fg / mL, the linear range is 10 fg / mL-50 ng / mL, and the method has an obvious potential application value in the clinical detection of procalcitonin.

Description

technical field [0001] The invention belongs to the field of novel nanometer materials and the technical field of biosensing. Background technique [0002] Ferritin is a kind of soluble protein that stores iron widely in animals, plants and microorganisms. It is a highly conserved biological molecule in evolution. It is abundant in the liver and spleen of mammals. The molecular structure of ferritin includes two parts: Protein shell and iron core. The protein shell surface of ferritin exposes many functional groups such as amino groups and hydroxyl groups, while the iron core part has been proven to catalyze the decomposition of hydrogen peroxide into superoxide radicals. Therefore, ferritin is not only easy to carry out surface chemical modification and It has a good peroxidase-like effect; N-(4-aminobutyl)-N-ethyl isoluminol, referred to as isoluminol, has excellent electrochemiluminescence properties and is used as a Derivatives, because the molecular structure contains ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N27/30G01N27/327
CPCG01N21/76G01N27/308G01N27/3278
Inventor 魏琴杨磊冯锐刘蕾刘雪静魏东闫涛
Owner UNIV OF JINAN
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