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based on hemin@fe 3 o 4 Chemiluminescent Immunodetection of Chicken Cytokines Using Mps Mimic Enzymes

A chemiluminescence immunoassay and cytokine technology, which is applied in the direction of measuring devices, scientific instruments, instruments, etc., can solve the problems that there are no multiple avian cytokines, joint detection of multi-component immunoassay methods, etc., and achieve easy operation and low cost , the effect of less consumption

Active Publication Date: 2019-08-09
YANGZHOU UNIV
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  • Claims
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AI Technical Summary

Problems solved by technology

At present, a variety of immunoassay methods have been developed to detect a single avian cytokine, but there is no report on a multi-component immunoassay method for the joint detection of multiple avian cytokines

Method used

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  • based on hemin@fe <sub>3</sub> o <sub>4</sub> Chemiluminescent Immunodetection of Chicken Cytokines Using Mps Mimic Enzymes
  • based on hemin@fe <sub>3</sub> o <sub>4</sub> Chemiluminescent Immunodetection of Chicken Cytokines Using Mps Mimic Enzymes
  • based on hemin@fe <sub>3</sub> o <sub>4</sub> Chemiluminescent Immunodetection of Chicken Cytokines Using Mps Mimic Enzymes

Examples

Experimental program
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Embodiment 1

[0023] The immunosensing array of this embodiment includes 4 rows×12 columns, a total of 48 detection sites, and each site has a diameter of 2 mm. ChIL-4, ChIFN-γ capture antibodies were immobilized in different rows. Each column can be used to detect 2 kinds of antigens in a single sample at the same time, so 12 columns can detect 2 kinds of cytokines contained in 12 samples at the same time, and can detect 24 samples at the same time. Based on Hemin@Fe 3 o 4 The method for the chemiluminescent immunodetection of chicken cytokines by MPs imitating enzymes, the specific steps are:

[0024] (1) Put the glass slide in piranha solution (H 2 SO 4 / 30%H 2 o 2 , 7:3 volume ratio) activated for 10-16 hours to make the surface with hydroxyl groups, rinsed with water and dried with nitrogen, then silanized with 1% GPTMS / toluene solution at room temperature overnight. Rinse with toluene and ethanol in turn to remove physically adsorbed silane, and blow dry with nitrogen after was...

Embodiment 2

[0029] 1. Make a standard curve of the linear relationship between the intensity of the chemiluminescent signal and the concentration of chicken cytokines

[0030] (1) Add 5 μL of antigen standard samples of different concentrations of ChIL-4 and ChIFN-γ to corresponding rows of different detection sites in the immunosensing array prepared in Example 1, incubate online for 20-30 min, wash and dry.

[0031] (2) Aminated Fe 3 o 4 Nanoparticles are synthesized by the classic hydrothermal method (C.Bendicho, F.Pena, M.Costas, et al.Photochemistry-based sample treatments as greenet approaches for trace-element analysis and specification.Trends Anal.Chem,29(2010) :681-691), hemin (Hemin) and aminated Fe 3 o 4 MPs were mixed at a molar ratio of 1:11, and the mixed solution was gently stirred for 0.5–1 h to form Hemin@Fe 3 o 4 MPs complex. Add 10 μL of 100 μg / mL ChIL-4 labeled antibody (ChIL-4-Ab 2 ) or ChIFN-γ-labeled antibody (ChIFN-γ-Ab 2 ), the mixed solution was gently st...

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Abstract

The invention discloses a Hemin@Fe based 3 o 4 Chemiluminescent Immunodetection of Chicken Cytokines Using MPs-mimicking Enzymes. The method adopts double-enzyme synergistic catalytic signal amplification technology, and the immunosensing array is made of silanized disposable glass slides, and the capture antibodies of different chicken cytokines are coated on the corresponding binding sites in a covalent manner, And the labeled antibodies of different chicken cytokines were immobilized on Hemin@Fe 3 o 4 Formation of corresponding Hemin@Fe on MPs nanoparticles 3 o 4 MPs-Ab 2 Signal amplifying nanoprobes. The invention forms a stable sandwich chicken cytokine immune complex through the specific recognition of the antigen by the secondary antibody, catalyzes the chemiluminescence reaction, and generates strong chemiluminescence. The detection range of the method of the invention is 0.005-0.1 ng / mL, and the chemiluminescence immunodetection of multi-component chicken cytokines with dual-enzyme synergistic catalytic signal amplification can be realized.

Description

technical field [0001] The invention belongs to the field of poultry cytokine detection, and specifically relates to a Hemin@Fe-based 3 o 4 Chemiluminescent Immunodetection of Chicken Cytokines Using MPs-mimicking Enzymes. Background technique [0002] Cytokines play an important role in body responses such as intercellular information transmission, body immunity, stimulation of hematopoietic stem cell regeneration, and participation in tissue repair through autocrine, paracrine, or endocrine methods. At present, there are few detection methods for cytokines, and the traditional detection methods are mostly biological and immunological methods, mainly including biological activity assays, immunoassays, and molecular biology methods. [0003] Zhao et al studied the effect of inflammation on acute myocardial ischemia-reperfusion in rabbits by performing enzyme-linked immunosorbent assay on interleukin-6 in rabbits and measuring the expression of IFN-γ in normal and infarcted...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/535G01N33/532G01N33/543
CPCG01N33/532G01N33/535G01N33/5436
Inventor 杨占军黄英李娟
Owner YANGZHOU UNIV
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