Microfluidics immune chip analysis method based on magnetic particle nano-enzyme, and application

A technology of microfluidic chip and immune chip, which is applied in the direction of analyzing materials, chemical instruments and methods, and material analysis through optical means, and can solve the problems of poor stability and low sensitivity of biological enzymes

Inactive Publication Date: 2019-01-18
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the microfluidic immune chip does not essentially solve the problems of low sensitivity and poor stability of biological enzymes in traditional enzyme-linked immunoassays.

Method used

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  • Microfluidics immune chip analysis method based on magnetic particle nano-enzyme, and application
  • Microfluidics immune chip analysis method based on magnetic particle nano-enzyme, and application
  • Microfluidics immune chip analysis method based on magnetic particle nano-enzyme, and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0108] This example is used to illustrate the preparation method of the immune chip of the present invention.

[0109] (1) Take the first microfluidic chip and seal it with the substrate, make the surface of channel I closely adhere to the substrate, pass the coating material into channel I to coat the substrate, and remove the first chip after the coating is completed Microfluidic Chip;

[0110] (2) Take the second microfluidic chip and seal it with the substrate, so that the surface of channel II is closely attached to the substrate and the arrangement direction of channel II is perpendicular to the direction of channel I, and the analyte is passed into channel II for incubation. followed by washing with buffer;

[0111] (3) Pass into channel II the secondary antibody labeled with magnetic particle nanozyme, wash with buffer after incubation, and the secondary antibody is an antibody against the analyte;

[0112] (4) Remove the second microfluidic chip, add a chemical reac...

Embodiment 2

[0118] This example is used to illustrate the steps of coupling morphine, cocaine and methamphetamine antibodies with magnetic particle nanozymes.

[0119] The three antibodies were purified by ultrafiltration through a 10Kd ultrafiltration tube, and then dissolved in PBS (pH7.2-7.4) buffer. First, add 10 μL, 20 mg / mL EDC and NHS coupling reagent to 100 μL, 5 mg / mL magnetic particle nanozyme, then react for 20 minutes, magnetically separate, resuspend with PBS solution, and then add 100 μL 1 mg / mL antibody, React at room temperature for 2 hours, then magnetically separate to remove excess antibody, and then store the magnetic particle-antibody conjugate in a 4-degree refrigerator.

Embodiment 3

[0120] The detection of embodiment 3 cocaine

[0121] This example is used to illustrate the application of the microfluidic detection chip of the present invention in the detection of cocaine.

[0122] The operation steps of the microfluidic chip detecting cocaine are as follows:

[0123] (1) Cocaine antigen stock dilution and coating: take 1 μL of cocaine antigen and add it to PBS (pH7.2-7.4) buffer solution, dilute to the optimal coating concentration, and mix well; take a multi-channel chip and seal it with the substrate; The diluted protein solution was passed into the three pipelines through the sample hole, and was coated for 30 minutes at room temperature.

[0124] (2) Blocking: 3% BSA blocking solution was prepared with PBS buffer. Remove the first chip, after air drying, paste the second chip, make the pipeline on the second chip perpendicular to the pipeline placement direction of the first chip, and seal; then use a pipette to inject 20 μL BSA into the pipeline...

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Abstract

The invention provides a microfluidics immune chip analysis method based on magnetic particle nano-enzyme, and application. The fermentoid activity of the magnetic particle nano-enzyme is used and serves as the substitute goods of HRP (horseradish peroxidase), and the stability of a traditional microfluidics immune chip can be greatly improved. In addition, the method is combined with high-sensitivity chemiluminescent immunoassay, and therefore, detection sensitivity can be greatly improved. Meanwhile, the microfluidics chip has the advantages that a sample amount is small, multiple samples and indexes can be simultaneously and quantitatively detected, and the microfluidics chip is high in specificity and sensitivity, low in price, quick, visual and convenient in operation, a reading result is quantifiable, complex instrument equipment and professional operation personnel are not required, and the microfluidics chip is suitable for quick field detection. Therefore, a powerful tool is provided for detecting micromolecules and biomacromolecules.

Description

technical field [0001] The invention belongs to the technical field of immune detection and analysis, in particular to a microfluidic immune chip analysis method based on a magnetic particle nano-enzyme signal readout system and its application. Background technique [0002] The construction of simple, stable and sensitive analytical methods has always been one of the goals pursued by the field of analytical chemistry. Simple, stable and sensitive analytical methods have promoted the development of analytical chemistry and its applications in clinical diagnosis, environmental monitoring, food safety, biological imaging and other fields. Simple and rapid analytical methods are particularly important in the early diagnosis of major infectious diseases, bacterial or viral infections, cancer and other diseases, as well as the screening of trace toxic substances in food and the environment. The immunological biomarker analysis method based on antibody-antigen specific recognitio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543G01N33/53G01N21/76B01L3/00
CPCB01L3/502707B01L2200/10G01N21/763G01N33/5304G01N33/535G01N33/54326G01N33/54346G01N2446/84G01N2446/90
Inventor 蒋兴宇阎锡蕴陈翊平段德民
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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