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Microfluidic immune chip analysis method based on biotin and streptavidine system

A streptavidin and microfluidic chip technology, which is applied in the direction of chemical reaction of materials for analysis, material analysis, biological testing, etc., to achieve the effect of small sample size, low price, and quantifiable reading results

Inactive Publication Date: 2016-02-17
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

[0007] The purpose of the present invention is to provide a microfluidic system based on biotin and streptavidin in view of the fact that the current immunoassay method cannot meet the actual application requirements in terms of detection sensitivity, analysis speed and signal readout mode. Immunochip Analysis Method and Its Application

Method used

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  • Microfluidic immune chip analysis method based on biotin and streptavidine system
  • Microfluidic immune chip analysis method based on biotin and streptavidine system
  • Microfluidic immune chip analysis method based on biotin and streptavidine system

Examples

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Effect test

Embodiment 1

[0071] Example 1 Morphine, cocaine and methamphetamine antibody biotinylation

[0072] The three antibodies were purified by ultrafiltration through a 10Kd ultrafiltration tube, then dissolved in PBS (pH7.2-7.4) buffer, mixed according to the molar ratio of antibody to biotin of 1:20, and the reaction was shaken on a vortex shaker 1h, then ultrafiltration through a 10Kd ultrafiltration tube to separate excess biotin, 10000r, 10min, then add 400uLPBS to wash, centrifuge 3 times, and finally store the biotinylated antibody at -20°C.

Embodiment 2

[0073] The detection of embodiment 2 cocaine

[0074] The application of the microfluidic detection chip of the present invention in the detection of cocaine.

[0075] The operation steps of the microfluidic chip detecting cocaine are as follows:

[0076] (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.

[0077] (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 pass 20 μL BSA into the pipeline to seal solution at room temperature ...

Embodiment 3

[0083] The detection of embodiment 3 morphine

[0084] The application of the microfluidic detection chip of the present invention in the detection of morphine.

[0085] The operation steps of the microfluidic chip detecting morphine are as follows:

[0086] (1) Dilution and coating of morphine antigen stock solution: Take 1 μL of morphine 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.

[0087] (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 pass 20 μL BSA into the pipeline to seal solution at ro...

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Abstract

The invention provides a microfluidic immune chip analysis method based on a biotin and streptavidine system. The method includes the following steps that firstly, a first microfluidic chip is acquired and sealed to a base, coverage substances are introduced into a channel for coverage, and the first chip is removed; secondly, a second microfluidic chip is taken and sealed to the base, and to-be-detected substances are introduced and are washed through buffering liquid after incubation; thirdly, a second antibody marked by biotin or streptavidine is introduced and is washed through a buffering liquid after incubation, and the second antibody is an antibody resistant to the to-be-detected substances; fourthly, HRP marked by streptavidine or biotin is introduced and is washed through a buffering liquid after incubation; fifthly, the second microfluidic chip is removed, HRP is added into a reaction area to catalyze light-emitting optical reaction liquid, and data are read through a chemical light-emitting instrument. The immune analysis method has the advantages that sensitivity is high, the signal reading mode is simple, the detection linearity range is wide, analysis speed is high, detection cost is low, and high-throughout test can be achieved.

Description

technical field [0001] The invention belongs to the technical field of immune detection and analysis, and in particular relates to a microfluidic immune chip analysis method based on a biotin and streptavidin signal amplification system and an application thereof. Background technique [0002] The construction of highly sensitive analytical methods has always been one of the goals pursued in the field of analytical chemistry. Highly 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. Highly sensitive 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 recognition is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/536G01N33/53G01N21/76
CPCG01N33/536G01N21/763G01N33/5302G01N33/581
Inventor 蒋兴宇陈翊平吴景
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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