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A salivary exosomal protein biomarker

A technology of salivary exosomes and biomarkers, which is applied in the field of salivary exosome protein biomarkers, can solve the problem of AD diagnostic methods without sensitive and effective non-invasive biomarkers

Active Publication Date: 2022-07-26
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 protein biomarker, which aims to solve the problem that the existing AD diagnostic methods have no sensitive, effective and non-invasive biomarkers

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0039] (1) Salivary exosome protein isolation

[0040] 1. Take a saliva sample and centrifuge at 2600 x 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 in a ratio of 2:1 to the sample volume, and vortex to mix evenly , incubate the centrifuge tube at 4°C overnight;

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

[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) , Pepstatin A), repeated suction...

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Abstract

The present invention provides a salivary exosome protein biomarker, wherein the salivary exosome protein biomarker is keratin, actin, histone, mitogen-activated protein kinase 1, calpain 1, myelin One or more of peroxidase, matrix metalloproteinase-9, and inflammatory protein. Keratin, actin, histone, mitogen-activated protein kinase 1, calpain 1, myeloperoxidase, matrix metalloproteinase-9, and inflammatory proteins of the present invention can be used as biomarkers for AD. Keratin, actin, histone, mitogen-activated protein kinase 1, calpain 1, myeloperoxidase, matrix metalloproteinase-9, inflammatory protein is a sensitive, effective and non-invasive salivary exosome Protein biomarkers, the use of the above salivary exosome protein biomarkers can detect and predict whether AD is present, so as to evaluate AD prevention, delay and drug intervention.

Description

technical field [0001] The invention relates to the technical field of biomarkers, in particular to a salivary exosome protein biomarker. Background technique [0002] Exosomes are nanoscale lipid inclusion structures with a diameter of 30-150 nm, which are cup-shaped under transmission electron microscopy and have a density range of 1.10-1.19 g / ml. They exist in a variety of 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 complexes, after the fusion of early endosomes, Exosomes are produced by the extracellular release. The production of exosomes can be caused by many factors, including extracellular stimuli, such as microbial attack and other stressful conditions. . In addition to the function of exosomes in the normal develop...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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