Biomarker of saliva exosom protein

A salivary exosome and biomarker technology, applied in the field of salivary exosome protein biomarkers, can solve the problems of insensitive, effective and non-invasive biomarkers in AD diagnosis methods

Active Publication Date: 2019-10-08
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the above deficiencies in the prior art, the purpose of the present invention is to provide a salivary exosome protei...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0039] (1) Isolation of salivary exosome protein

[0040] 1. Take a saliva sample and centrifuge at 2600×g for 30 minutes at 4°C to remove cells, bacteria, debris and food residues;

[0041] 2. Take the saliva supernatant, centrifuge at 14,000×g for 2 minutes at room temperature, transfer the supernatant to a new centrifuge tube, add exosome precipitation reagent at a ratio of 2:1 to the sample volume, and vortex to mix evenly , incubate the centrifuge tube overnight at 4°C;

[0042] 3. After incubation, centrifuge at 10,000×g for 1 hour at 4°C, discard the supernatant, and centrifuge at 14,000×g for 10 seconds at room temperature to remove the liquid attached to the tube wall and obtain exosome pellets.

[0043] (2) Salivary exosome protein extraction

[0044] Resuspend the exosome pellet in urea buffer (50mM Tris-Hcl, 8M urea, 2M thiourea, 1mM DTT, pH=7.5), add protease inhibitor cocktail (AEBSF, Aprotinin, Bestatin, E-64, Leupeptin , PepstatinA), repeated suction and mixin...

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PUM

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Abstract

The invention provides a biomarker of saliva exosom protein. The biomarker includes one or multiple selected from keratin, actin, histone, mitogen activated protein kinase 1, calpain 1, myeloperoxidase, matrix metalloprotease-9 and inflammation protein. The above can serves as a biomarker of AD, and is sensitive, effective and atraumatic. The biomarker can detect and predict whether one has AD, ADcan be prevented and alleviated, and the drug intervention condition can be evaluated.

Description

technical field [0001] The invention relates to the technical field of biomarkers, in particular to a salivary exosome protein biomarker. Background technique [0002] Exosome is a nano-scale lipid inclusion structure with a diameter of 30-150 nm. It is cup-shaped under a transmission electron microscope and has a density in the range of 1.10-1.19 g / ml. It exists in various body fluids, such as serum, Saliva, cerebrospinal fluid, semen and urine. Exosomes originate from vesicles in the late endosome lumen, the cell membrane invaginates to form early endosomes, and then endocytoses related proteins. Under the regulation of Ras superfamily GTPase Rab and lipid raft complex, after early inner membrane fusion, released to the outside of the cell to produce exosomes. The production of exosomes can be caused by many factors, including extracellular stimuli, such as microbial attack and other stress conditions, etc. The release of exosomes is an effective regulation of cells to r...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/58G01N30/02G01N30/72G01N30/88
CPCG01N33/6848G01N33/583G01N33/6896G01N30/02G01N30/724G01N30/88G01N2458/00G01N2800/2821G01N2030/027G01N2030/8813
Inventor 王勇曾琼兰程思明倪嘉缵
Owner SHENZHEN UNIV
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