Method for extracting exosome in urine and method for analyzing proteomics and phosphorylation proteomics of exosome

Abstract

The invention discloses a method for extracting an exosome in urine and a method for analyzing proteomics and phosphorylation proteomics of the exosome. The invention firstly provides a magnetic nanomaterial with metal ions chelated on the surface. The magnetic nano material is prepared according to the steps of modifying magnetic nano particles on the surface of graphene oxide to obtain a product 1; coating the surface of the product 1 with a silicon dioxide layer to obtain a product 2; modifying the surface of the silicon dioxide layer of the product 2 with a silane coupling agent with amino to obtain a product 3; modifying the surface of the product 3 with a metal chelating agent to obtain a product 4; and chelating metal ions on the surface of the product 4. The magnetic nano materialcan be used for extracting the exosome in urine, and can be used for proteomics analysis or phosphorylation proteomics analysis of the exosome. According to the invention, phosphate radicals exposedon the surface of a membrane phospholipid bilayer of the exosome are combined with the metal ions through a double-coordination chemical effect, the high-specificity extraction of the urine exosome isrealized by combining an electrostatic adsorption effect at the same time, the extraction speed is high, and the recovery rate is high.

Description

technical field

[0001] The invention relates to the extraction of exosomes in urine and its proteomics and phosphorylated proteomics analysis methods, and relates to the fields of biotechnology and analytical chemistry. Background technique

[0002] Phosphorylation is an important post-translational modification of proteins, and phosphorylated proteins are involved in many important physiological processes, such as cell apoptosis, cell proliferation, and cell migration. Changes in the phosphorylation level of peptides are closely related to the occurrence and development of many diseases, and have potential application prospects as disease biomarkers. However, due to the unstable state of protein phosphorylation and the high abundance of phosphatases in body fluids, it brings great difficulties to the detection of phosphorylated proteins and phosphorylated peptides, thus limiting the use of phosphorylated proteins in the early diagnosis of diseases. Applications. Exosomes ...

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