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A ferritin-based encapsulation ir(ppy) 3 The preparation method of the biosensor

A technology of biosensor and ferritin, which is applied in the field of new nanomaterials and biosensing, can solve the problems of high detection cost and achieve the effect of good reproducibility, high sensitivity and simple preparation

Active Publication Date: 2021-10-26
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the concentration of tryptase is mainly detected by some expensive large-scale chromatography, atomic emission spectrometry and other instruments, which often require professionals to detect and the detection cost is expensive. hotspot

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0024] Embodiment 1. A kind of encapsulation Ir (ppy) based on ferritin 3 A biosensor preparation method 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) Immerse the glassy carbon electrode in 1% chloroauric acid solution to electrodeposit a layer of nano-gold as the sensing substrate, and drop 6 μL of capture antibody Ab at a concentration of 10 ug / mL on the surface 1 solution, wash the electrode surface with pH 7.4 phosphate buffer solution PBS, and incubate it at 4 °C for 12 h;

[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 tryptase standard solution or tryptase solution of ...

Embodiment 2

[0030] Embodiment 2. A kind of encapsulation Ir(ppy) based on ferritin 3 A biosensor preparation method comprising the following steps:

[0031] (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;

[0032] (2) Immerse the glassy carbon electrode in 2% chloroauric acid solution to electrodeposit a layer of nano-gold as the sensing substrate, and drop 6 μL of capture antibody Ab at a concentration of 10 ug / mL on the surface 1 solution, wash the electrode surface with pH 7.4 phosphate buffer solution PBS, and incubate it at 4 °C for 12 h;

[0033] (3) Add 3 μL of 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;

[0034](4) Add 6 μL tryptase standard solution or tryptase solution of...

Embodiment 3

[0036] Embodiment 3. A kind of encapsulation Ir(ppy) based on ferritin 3 A biosensor preparation method comprising the following steps:

[0037] (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;

[0038] (2) Immerse the glassy carbon electrode in 3% chloroauric acid solution to electrodeposit a layer of nano-gold as the sensing substrate, and add 6 μL of capture antibody Ab at a concentration of 10 ug / mL on the surface 1 solution, wash the electrode surface with pH 7.4 phosphate buffer solution PBS, and incubate it at 4 °C for 12 h;

[0039] (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;

[0040] (4) Add 6 μL tryptase standard solution or tryptase solution of un...

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Abstract

A ferritin-based encapsulation of Ir(ppy) 3 The method for preparing a biosensor belongs to the field of novel nanomaterials and the field of biosensing technology; the present invention utilizes a pH-guided protein depolymerization / recombination method to encapsulate a large amount of three (2-phenylpyridine) iridium in apoferritin Ft Ir(ppy) 3 The molecule gets Ft‑Ir(ppy) 3 As the energy donor of electrochemiluminescence ECL, the glassy carbon electrode surface is modified with nano gold as the ECL energy acceptor, based on Ir(ppy) for the first time 3 Excellent ECL performance and its principle of ECL resonance energy transfer with gold nanoparticles proposed a biosensor preparation method with simple preparation, low cost, low reaction energy consumption, green environmental protection and high sensitivity, and applied it to tryptase For actual sample detection, the detection limit is as low as 1.3 fg / mL, and the linear range is as wide as 5 fg / mL-100 ng / mL. It has high sensitivity and good reproducibility, and has great potential application value.

Description

technical field [0001] The invention belongs to the field of novel nanometer materials and the technical field of biosensing. Background technique [0002] As a research hotspot arising from the intersection of multiple disciplines such as biology, chemistry, medicine, and electronic technology, electrochemiluminescence ECL immunoassay technology is an organic combination of electrochemistry, chemiluminescence, and immunoassay technologies. With the advantages of good selectivity, high sensitivity, fast analysis speed, easy automation, miniaturization and integration, etc., it has been widely used in the fields of disease marker analysis, food safety analysis, environmental pollution analysis and other fields. [0003] Tryptase is a specific protease produced by the human body when allergic reactions occur. It can be used as an indicator of allergic reactions. Its sensitive detection can provide effective early warning for human allergic reactions, control the disease in tim...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/76G01N27/30G01N27/327
CPCG01N21/76G01N27/308G01N27/3278
Inventor 杨磊魏琴范大伟冯瑞卿王欢张诺任祥
Owner UNIV OF JINAN
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