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Rapid detection platform and method ofPD-L1 exosome in vitro

A technology for PD-L1 and in vitro detection, applied in the direction of measuring devices, biological tests, material inspection products, etc., can solve the problems of inconvenient operation, high cost, and low specificity of exosome detection, and achieve convenient operation, low cost, Check the effect of shortcut

Inactive Publication Date: 2019-06-14
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The invention solves the technical problems of low specificity, inconvenient operation and high cost of exosome detection in the prior art

Method used

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  • Rapid detection platform and method ofPD-L1 exosome in vitro
  • Rapid detection platform and method ofPD-L1 exosome in vitro

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] In vitro rapid detection platform and method for PD-L1 exosomes, including:

[0031] Sample primary filtration area 1: The sample primary filtration area is glass cellulose for primary filtration of whole blood, retaining most of the blood cells;

[0032] Nanofiltration area 2: The nanofiltration area 2 is connected to the sample primary filtration area 1; the nanofiltration area 2 is a nano-hydrophilic filter membrane, which realizes the further interception of residual blood cells in the previous step and at the same time realizes the separation of exosomes; nanofiltration The diameter of the membrane is 150 nm.

[0033] Chromogenic material pad area 3: the chromogenic material pad area 3 is connected to the nanofiltration area 2; the chromogenic material pad area 3 is used to fix the chromogenic material-labeled antibody, and can make the chromogenic material-labeled antibody and the sample complex dissolved; the chromogenic substance-labeled antibody is used to spe...

Embodiment 2

[0038] In vitro rapid detection platform and method for PD-L1 exosomes, including:

[0039] Sample primary filtration area 1: The sample primary filtration area is glass cellulose for primary filtration of whole blood, retaining most of the blood cells;

[0040]Nanofiltration area 2: The nanofiltration area 2 is connected to the sample primary filtration area 1; the nanofiltration area 2 is a nano-hydrophilic filter membrane, which realizes the further interception of residual blood cells in the previous step and at the same time realizes the separation of exosomes; nanofiltration The diameter of the membrane is 220 nm.

[0041] Chromogenic material pad area 3: the chromogenic material pad area 3 is connected to the nanofiltration area 2; the chromogenic material pad area 3 is used to fix the chromogenic material-labeled antibody, and can make the chromogenic material-labeled antibody and the sample complex dissolved; the chromogenic substance-labeled antibody is used to spec...

Embodiment 3

[0045] In vitro rapid detection platform and method for PD-L1 exosomes, including:

[0046] Sample primary filtration area 1: The sample primary filtration area is glass cellulose for primary filtration of whole blood, retaining most of the blood cells;

[0047] Nanofiltration area 2: The nanofiltration area 2 is connected to the sample primary filtration area 1; the nanofiltration area 2 is a nano-hydrophilic filter membrane, which realizes the further interception of residual blood cells in the previous step and at the same time realizes the separation of exosomes; nanofiltration The diameter of the membrane is 250 nm.

[0048] Chromogenic material pad area 3: the chromogenic material pad area 3 is connected to the nanofiltration area 2; the chromogenic material pad area 3 is used to fix the chromogenic material-labeled antibody, and can make the chromogenic material-labeled antibody and the sample complex dissolved; the chromogenic substance-labeled antibody is used to spe...

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Abstract

The invention discloses a rapid detection platform and method of PD-L1 exosome in vitro, and belongs to the technical field of rapid detection. The detection platform comprises a sample initial-filtration area, a nanofiltration area, a coloration object pad area, a coloration area and a water absorption area. In a capture antibody on the coloration area and a coloration object labeled antibody onthe coloration object pad area, at least one is an exosomePD-L1 antibody. The detection method comprises the steps that a to-be-tested sample is filtered initially through an initial-filtering film, then the to-be-tested sample is filtered through a nanofiltration film, and thus the exosome is obtained; the exosome is specifically combined with the coloration object labeled antibody and then combined with the capture antibody to form a double-antibody sandwich; and in the capture antibody and the coloration object labeled antibody, at least one is the exosome PD-L1 antibody. According to the rapid detection platform of the PD-L1 exosome in vitro, whole blood separation, exosome purifying and special detection of the PD-L1exosome are integrated by the platform, and the characteristics thatthe operation is convenient, detection is fast, and the cost is low are achieved.

Description

technical field [0001] The invention belongs to the technical field of rapid detection, and in particular relates to an in vitro rapid detection platform and detection method of PD-L1 exosomes. Background technique [0002] Exosomes are nanoscale vesicles (50-150nm in diameter) secreted by cells into the extracellular microenvironment, which were first discovered in 1983. However, due to the lack of analytical means, exosomes have long been considered as a tool for cells to excrete intracellular garbage. With the continuous innovation of analytical methods, the mechanism of exosome secretion has been gradually revealed. Now, the mainstream view in the scientific community is that exosomes are mainly involved in intercellular signal transmission and as disease markers. The 2013 Nobel Prize in Physiology or Medicine was awarded to three outstanding scientists who revealed major discoveries in this field. However, the resulting exosome function research and potential future ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/533G01N33/535G01N21/78G01N21/64G01N1/34
Inventor 刘笔锋陈鹏
Owner HUAZHONG UNIV OF SCI & TECH
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