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Preparation method of Au@Fe3O4MNPs-Ab2 nano enzyme detection probe and method for detecting multi-component antigen

A technology for detecting probes and nanozymes, applied in nanotechnology, measuring devices, analytical materials, etc., can solve problems such as limited catalytic activity, and achieve the effects of improving detection throughput, low cost, and reducing detection costs

Active Publication Date: 2019-04-26
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The present invention aims at the problem of limited catalytic activity of nanomaterial amplification carriers used in chemiluminescent immunoassays in the prior art to improve analytical sensitivity, and provides an Au@Fe 3 o 4 MNPs-Ab 2 The preparation method of nanozyme detection probe, to make a kind of Au and diFe 3 o 4 The nanozyme detection probe of conjoined structure makes Fe 3 o 4 The catalytic activity is not blocked, improving the detection sensitivity

Method used

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  • Preparation method of Au@Fe3O4MNPs-Ab2 nano enzyme detection probe and method for detecting multi-component antigen
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  • Preparation method of Au@Fe3O4MNPs-Ab2 nano enzyme detection probe and method for detecting multi-component antigen

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Embodiment 1

[0028] A kind of Au@Fe is prepared through this embodiment 3 o 4 MNPs-Ab 2 Nanozyme detection probe, the specific process is:

[0029] Firstly, Au nanoparticles were prepared: 1.0 g HAuCl4 3H2O (2.5 mmol) and 10 mL oleylamine (30 mmol) were sequentially added to 100 mL tetralin, heated at 65 °C for 5 h, cooled to room temperature, collected by centrifugation and used Au nanoparticles were obtained by washing with ethanol, which were dispersed in hexane to obtain a dispersion colloid of Au nanoparticles with a dispersion concentration of 10 mg / mL.

[0030] Re-preparation of Au@Fe 3 o 4 Nanoparticles: Mix 1 mL of oleic acid (3 mmol) with 20 mL of octadecene in N 2 Heating under flow to 120°C for 20 min, under N 2 layer, inject 0.15 mL of Fe(CO) at a concentration of 5 mg / mL into the solution 5 . After stirring for 5 min, 0.5 mL of oleylamine was injected into the reaction mixture, and then 2 mL of the aforementioned colloid of Au nanoparticles was injected. Heat the sol...

Embodiment 2

[0034] In this example, the Au@Fe prepared in Example 1 3 o 4 MNPs-Ab 2 A method for detecting multi-component antigens by a chemiluminescent array immunosensor prepared from a nanozyme detection probe, comprising the following steps:

[0035] B1: Immerse the disposable glass slide in H at a volume ratio of 7:3 2 SO 4 and 30%H 2 o 2 Activate in piranha acid solution for 10-12 hours to make the surface with amino groups, rinse with deionized water and dry, then soak in 1% GPTMS (γ-glycidyl ether oxypropyltrimethoxysilane) Silanize it in a toluene solution, rinse it with toluene and ethanol in turn, and dry it with nitrogen to make a silanized glass slide; then use screen printing technology and use a template to print the silanized glass slide into 4 rows×12 A reaction array with 48 detection sites in a column format, in which the holes formed in the hydrophobic non-photoactive membrane have a diameter of 2 mm, and the distance between the holes on the edge and the edge o...

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Abstract

The invention relates to a preparation method of an Au@Fe3O4MNPs-Ab2 nano enzyme detection probe and a method for detecting a multi-component antigen. Firstly, compounding an Au@Fe3O4 nano particle probe, and successfully applying the Au@Fe3O4 nano particle probe to construct a chemiluminiscence array immunosensor, secondly, fixing different capture antibodies on different solid phase interfaces of the array immunosensor, respectively introducing an antigen sample and an Au@Fe3O4MNPs-Ab2 nano signal amplification probe, and performing on-line incubation to form a stable sandwich immune reaction complex; wherein the light signal generated after the chemiluminescence substrate is introduced is collected by a charge coupled CCD camera, so that the chemiluminescence immune detection of the multi-component antigen amplified by the nano enzyme catalytic signal is achieved, and the method is suitable for the simultaneous detection of a plurality of analyte antigens. The analysis method has the advantages of low detection cost, low sample consumption, short consumption time, good stability, simple operation and the like, and provides a detection platform with a deep prospect for clinical detection of poultry diseases.

Description

technical field [0001] The invention relates to the technical field of biomolecular chemiluminescence immunoassay detection and analysis, in particular to an Au@Fe 3 o 4 MNPs-Ab 2 A preparation method of a nanozyme detection probe and a method for detecting multi-component antigens by a luminescence array. Background technique [0002] Chemiluminescence Immunoassay (CLIA) is an analysis method combining high-sensitivity chemiluminescence and high-specificity immune response. A new type of labeled immunoassay technology for detecting trace antigens and antibodies developed after immunoassay methods such as immunoassay. Chemiluminescence imaging immunoassay is one of the most concerned methods in chemiluminescence immunoassay detection at present. It combines chemiluminescence immunoassay and imaging technology, and has the advantages of chemiluminescence immunoassay and high throughput of imaging analysis, and can multi-component Advantages of simultaneous detection. It ...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/531G01N21/76B82Y40/00
CPCG01N21/76G01N33/531G01N33/543B82Y40/00
Inventor 胡锐宣刘靳一蒙吴昕玥杨占军李娟
Owner YANGZHOU UNIV
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