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Glycosylation research method of exosomes

An exosome and glycosylation technology, applied in biochemical equipment and methods, epidermal cells/skin cells, material separation, etc., can solve problems such as inappropriate exosome glycosylation function research, and maintain integrity and activity, small influence, small effect of steric hindrance

Active Publication Date: 2021-08-13
XIAMEN UNIV
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Problems solved by technology

[0007] The above patents are not directly related to the glycosylation of exosomes, therefore, they are also not applicable to the research on the glycosylation function of exosomes

Method used

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  • Glycosylation research method of exosomes
  • Glycosylation research method of exosomes
  • Glycosylation research method of exosomes

Examples

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Embodiment Construction

[0035] In the following examples, the study on the glycosylation of exosomes expressing PD-L1 is taken as an example.

[0036] see figure 1 , cells were treated with azide-modified sialic acid metabolic substrate Ac 4 ManNAz culture, the cells metabolize sugar to the surface of the cell membrane to form azide-modified sialic acid, and then secrete exosomes through endocytosis, so that the azide-modified sialic acid is labeled on the surface of the exosome membrane, followed by DBCO-Cy5 alkyne Copper-free bioorthogonal reaction was carried out to verify the labeling efficiency of glucose metabolism with base dye (dissolved in DMSO). (The exosomes of A375 cells cultured with an equal amount of DMSO were used as the control group).

[0037] (1) Cell culture. A375 cells (melanoma) were digested and resuspended in DMEM medium containing 1% FBS and 1% double antibody, and cultured overnight. After the A375 cells adhered to the wall, the DMEM medium was removed, and the DMEM medi...

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Abstract

The invention discloses a glycosylation research method of exosomes, which comprises the following steps of: 1) co-culturing cells secreting the exosomes and non-natural monosaccharides, wherein the cells secretes the exosomes containing the non-natural monosaccharides; (2) marking a functional probe on the non-natural sugar, wherein the functional probe is provided with a first mark; and 3) detecting the first marker to obtain exosome information. The method disclosed by the invention is simple to operate and does not need complex sample pretreatment; and the exosomes does not need to be cracked.

Description

technical field [0001] The invention relates to a method for studying the glycosylation of exosomes. Background technique [0002] The surface of exosomes is covered by glycoproteins, and the glycosylation of exosome proteins plays an important role in transmembrane signal transmission, regulation of functions and physiological patterns between exosomes and recipient cells, and tumor progression and metastasis. Therefore, studying the glycosylation of exosomes is helpful to understand the physiological functions involved in exosomes and to develop disease-related diagnostic and therapeutic markers. [0003] Although the importance of glycosylation in exosomes has been gradually recognized in physiological processes, the sugar metabolism mechanism of exosomes is unknown, the composition of sugar chains is complex, and the modification sites are diverse. Therefore, the characterization and function of glycosylation Research is very difficult. The main methods currently used ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/63G01N27/62G01N30/02C12N5/09
CPCG01N21/63G01N27/62G01N30/02C12N5/0693C12N5/0626C12N2500/34
Inventor 杨朝勇朱琳许元风林颢霆黄梦娇宋彦龄
Owner XIAMEN UNIV
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