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Biosensor preparation method based on ferritin-encapsulated Ir(ppy)3

A biosensor and ferritin technology, applied in chemiluminescence/bioluminescence, instruments, measuring devices, etc., can solve the problems of high detection costs and achieve good reproducibility, fast response, and low cost

Active Publication Date: 2019-11-12
UNIV OF JINAN
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  • Summary
  • Abstract
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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
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1. A kind of encapsulation Ir (ppy) based on ferritin 3 The biosensor preparation method is characterized in that, 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) Put the glassy carbon electrode into 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 of tryptase standard solu...

Embodiment 2

[0030] Embodiment 2. A kind of encapsulation Ir(ppy) based on ferritin 3 The biosensor preparation method is characterized in that, 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) Put the glassy carbon electrode into 2% chloroauric acid solution to electrodeposit a layer of nano-gold as the sensing substrate, and drop 6 μL of capture antibody Ab with 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 of tryptase standard s...

Embodiment 3

[0036] Embodiment 3. A kind of encapsulation Ir(ppy) based on ferritin 3 The biosensor preparation method is characterized in that, 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) Put the glassy carbon electrode into the 3% chloroauric acid solution to electrodeposit a layer of nano-gold as the sensing substrate, and drop 6 μL of capture antibody Ab with 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 of tryptase standard...

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Abstract

The present invention provides a biosensor preparation method based on ferritin-encapsulated Ir(ppy)3, and belongs to the technical field of novel nanomaterials and biosensing. According to the methodprovided by the present invention, a pH-guided protein depolymerization / recombination method is used to encapsulate a large number of tris(2-phenylpyridyl)firpic Ir(ppy)3 molecules in the apoferritinFt to obtain Ft-Ir(ppy)3 as an electrochemiluminescence ECL energy donor, the surface of the glassy carbon electrode modified with nanogold is taken as an ECL energy acceptor, and based on the excellent ECL performance of Ir(ppy)3 and the ECL resonance energy transfer principle with nanogold, a biosensor preparation method with a simple preparation, a low cost, low reaction energy consumption, green environmental protection and high sensitivity is proposed for the first time; the method is applied to the actual sample detection of tryptase, the detection limit is as low as 1.3 fg / mL, and thelinear range is as wide as 5 fg / mL-100 ng / mL; and the method has high sensitivity, good reproducibility, and large 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 Applications(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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