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Preparation method and application of immunosensor promoting gold nano-cluster electroluminescent response based on iron oxide array co-reaction

An immunosensor, gold nanocluster technology, applied in the direction of analysis, chemiluminescence/bioluminescence, instrument, etc. by chemical reaction of materials, can solve the problems of poor conductivity of bovine serum albumin, low ECL efficiency, etc., and achieve wide detection. Effects of range, enhanced ECL emission, low detection limit

Inactive Publication Date: 2019-12-10
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In particular, gold nanoclusters using bovine serum albumin as a template can not only be used as ECL signal labels for low-concentration detection of disease markers due to their good bioidentity, but also can pass through a large number of amino groups, carboxyl groups, etc. on the surface of bovine serum albumin. Functional groups link immune molecules; however, due to the poor conductivity and low ECL efficiency of bovine serum albumin, it is particularly important to find a strategy to amplify its ECL response to meet the detection of trace markers

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1. A method for preparing an immunosensor based on the co-reaction of iron oxide arrays to promote the electroluminescent response of gold nanoclusters, comprising the following steps:

[0030] (1) Ultrasonic cleaning of ITO glass with a size of 1×2 cm in acetone for 1 h, followed by cleaning with water and absolute ethanol, and drying with nitrogen;

[0031] (2) Put the treated ITO glass into the reactor, add 30 mL of raw material mixture, and react at 120 °C for 6 h;

[0032] (3) After cooling, take out the ITO glass, rinse it with ultrapure water and absolute ethanol, dry it, put it in a tube furnace, and calcinate it at 200 °C for 3 hours under a nitrogen atmosphere to obtain Fe 2 o 3 Arrays, as sensor substrate materials;

[0033] (4) in Fe 2 o 3 Drop-coat 6 μL of Ab at a concentration of 2 mg / mL on the surface 1 solution, rinse the ITO surface with PBS of pH 7.4 after drying at 4 °C;

[0034] (5) Add 3 μL bovine serum albumin solution with a mass ...

Embodiment 2

[0037] Embodiment 2. A method for preparing an immunosensor based on iron oxide array co-reaction to promote the electroluminescent response of gold nanoclusters, characterized in that it comprises the following steps:

[0038] (1) Ultrasonic cleaning of ITO glass with a size of 1×2 cm in acetone for 1 h, followed by cleaning with water and absolute ethanol, and drying with nitrogen;

[0039] (2) Put the treated ITO glass into the reactor, add 40 mL of raw material mixture, and react at 135 °C for 7 h;

[0040] (3) After cooling, take out the ITO glass, rinse it with ultrapure water and absolute ethanol, dry it, put it in a tube furnace, and calcinate it at 250 °C for 3 hours under a nitrogen atmosphere to obtain Fe 2 o 3 Arrays, as sensor substrate materials;

[0041](4) in Fe 2 o 3 Drop-coat 6 μL of Ab at a concentration of 3.5 mg / mL on the surface 1 solution, rinse the ITO surface with PBS of pH 7.4 after drying at 4 °C;

[0042] (5) Add 3 μL bovine serum albumin solu...

Embodiment 3

[0045] Embodiment 3. A method for preparing an immunosensor based on iron oxide array co-reaction to promote gold nanocluster electroluminescence response, characterized in that it comprises the following steps:

[0046] (1) Ultrasonic cleaning of ITO glass with a size of 1×2 cm in acetone for 1 h, followed by cleaning with water and absolute ethanol, and drying with nitrogen;

[0047] (2) Put the treated ITO glass into the reactor, add 50 mL of raw material mixture, and react at 150 °C for 8 hours;

[0048] (3) After cooling, take out the ITO glass, rinse it with ultrapure water and absolute ethanol, dry it, put it in a tube furnace, and calcinate it at 300 °C for 3 hours under a nitrogen atmosphere to obtain Fe 2 o 3 Arrays, as sensor substrate materials;

[0049] (4) in Fe 2 o 3 Drop-coat 6 μL of Ab at a concentration of 5 mg / mL on the surface 1 solution, rinse the ITO surface with PBS of pH 7.4 after drying at 4 °C;

[0050] (5) Add 3 μL bovine serum albumin solution...

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Abstract

The invention relates to a preparation method and application of an immunosensor promoting gold nano-cluster electroluminescent response based on iron oxide Fe2O3 array co-reaction, and belongs to thefield of electrochemistry and immunoassay. Based on an electroluminescent ECL technology; C-reactive protein CRP is used as a target analyte; a gold nanocluster AuNCs is used as an ECL emitter; underthe catalysis of a co-reaction accelerant ferric oxide nano array Fe2O3, ECL emission of the AuNCs can be effectively enhanced by all exposed active sites because the transport of electrons along anordered array structure is improved, and therefore, Fe2O3 / AuNCs is used as a sensing platform and the sandwich type ECL immunosensor is constructed. The immunosensor has the advantages that the immunosensor shows high sensitivity, a wide detection range and a low detection limit for the target CRP; the immunosensor can achieve rapid and sensitive detection of the CRP in human serum, and the immunosensor is high in application value for clinically judging infection types of patients.

Description

technical field [0001] The invention belongs to the field of electrochemistry and immune analysis. Background technique [0002] Gold nanoclusters, a traditional fluorescent probe, are promising electrochemiluminescent agents due to their high quantum yield and large Stokes shift. In particular, gold nanoclusters using bovine serum albumin as a template can not only be used as ECL signal labels for low-concentration detection of disease markers due to their good bioidentity, but also can pass through a large number of amino groups, carboxyl groups, etc. on the surface of bovine serum albumin. Functional groups link immune molecules; however, due to the poor conductivity and low ECL efficiency of bovine serum albumin, it is particularly important to find a strategy to amplify its ECL response to meet the detection of trace markers. [0003] So far, using S 2 o 8 2− There are many studies on co-reactants for ECL emission of semiconductor quantum dots, the reason is that S ...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N27/327G01N33/531
CPCG01N21/76G01N27/3275G01N27/3278G01N33/531
Inventor 魏琴贾越杨磊赵磊张诺马洪敏
Owner UNIV OF JINAN
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