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au@fe 3 o 4 mnps-ab 2 Preparation method of nanozyme detection probe and method for detecting multi-component antigen

A technology for detecting probes and nanozymes, which can be used in nanotechnology, measurement devices, analytical materials, etc., and can solve problems such as limited catalytic activity.

Active Publication Date: 2022-04-29
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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  • au@fe <sub>3</sub> o <sub>4</sub> mnps-ab <sub>2</sub> Preparation method of nanozyme detection probe and method for detecting multi-component antigen
  • au@fe <sub>3</sub> o <sub>4</sub> mnps-ab <sub>2</sub> Preparation method of nanozyme detection probe and method for detecting multi-component antigen
  • au@fe <sub>3</sub> o <sub>4</sub> mnps-ab <sub>2</sub> Preparation method of nanozyme detection probe and method for detecting multi-component antigen

Examples

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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 The Au nanoparticles were washed with ethanol, and dispersed in hexane to obtain a dispersion 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 dispersion of Au nanoparticles prepared above was injected, the solution was heated to reflux ...

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 present invention relates to a kind of Au@Fe 3 o 4 MNPs‑Ab 2 Preparation method of nanozyme detection probe and method for detection of multi-component antigen, Au@Fe synthesized first in the present invention 3 o 4 Nanoparticle probes were successfully applied to construct chemiluminescent array immunosensors for nanozyme signal amplification. Secondly, different capture antibodies were immobilized on different solid phase interfaces of the immunoarray sensor, and then passed through the antigen sample and Au@Fe respectively. 3 o 4 MNPs‑Ab 2 The nano-signal amplification probe is incubated online to form a stable sandwich immune reaction complex, and the light signal generated after passing through the chemiluminescent substrate is collected by a charge-coupled CCD camera, realizing the multi-component antigen chemistry of nanozyme-catalyzed signal amplification Luminescent immunoassay for simultaneous detection of multiple analyte antigens. The analysis method has the advantages of low detection cost, less sample consumption, short time-consuming, good stability, simple operation, etc., and provides a promising detection platform 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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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/531G01N21/76B82Y40/00
CPCG01N21/76G01N33/531G01N33/543B82Y40/00
Inventor 杨占军胡锐宣刘靳一蒙吴昕玥李娟
Owner YANGZHOU UNIV
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