Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A kind of phosphopeptide solid-phase extraction column and its preparation and application

A technology of solid-phase extraction cartridges and phosphopeptides, which is applied in chemical instruments and methods, other chemical processes, instruments, etc., can solve the problems of inapplicable analysis of trace samples, long time consumption, etc., to reduce the complexity of interference and improve the preparation method Simple, highly specific effects

Active Publication Date: 2021-02-26
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional immobilized titanium ion affinity chromatography material is monodisperse, and the particle size of the material is about 12 microns. Generally, the material is dispersed in the solution to enrich the phosphopeptide, but this enrichment method requires repeated incubation and centrifugation steps. , takes a long time and is not suitable for analyzing trace samples (document 4. Zhou, H.; Ye, M. et al. Robust phosphoproteome enrichment using monodisperse microsphere–based immobilized titanium (IV) ion affinity chromatography. Nat. Protoc. 8, 461-480 ( 2013).)

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of phosphopeptide solid-phase extraction column and its preparation and application
  • A kind of phosphopeptide solid-phase extraction column and its preparation and application
  • A kind of phosphopeptide solid-phase extraction column and its preparation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028]The phosphopeptide solid phase extraction cartridge is used for the enrichment of phosphopeptides in the enzymatic hydrolysate of the mixture of bovine serum albumin and β-casein:

[0029](1) Disperse 1 g of polyvinyl alcohol into 120 mL of deionized water to obtain a dispersion medium;

[0030](2) Mix 18 mL of toluene, 18 mL of ethylene glycol dimethacrylate, 20 mL of glycidyl methacrylate and 0.12 g of benzoyl peroxide to obtain a monomer phase;

[0031](3) Add the monomer phase of step (2) to the dispersion medium of step (1) and heat and stir at 70° C. for 12 hours to obtain polymer microspheres;

[0032](4) Reacting the polymer microspheres obtained in step (3) with 150 mL of ethylenediamine to obtain polymer microspheres with amino groups on the surface;

[0033](5) Reacting the polymer microspheres with amino groups on the surface obtained in step (4) with 5 mL phosphoric acid, and then reacting with 100 mM titanium sulfate at 25° C. for 16 hours to prepare a fixed titanium ion affini...

Embodiment 2

[0044]The phosphopeptide solid phase extraction cartridge is used for the enrichment of phosphopeptides in the HeLa hydrolysis solution:

[0045](1) Disperse 1 g of polyvinyl alcohol into 120 mL of deionized water to obtain a dispersion medium;

[0046](2) Mix 18 mL of toluene, 18 mL of ethylene glycol dimethacrylate, 20 mL of glycidyl methacrylate and 0.12 g of benzoyl peroxide to obtain a monomer phase;

[0047](3) Add the monomer phase of step (2) to the dispersion medium of step (1) and heat and stir at 70° C. for 12 hours to obtain polymer microspheres;

[0048](4) Reacting the polymer microspheres obtained in step (3) with 150 mL of ethylenediamine to obtain polymer microspheres with amino groups on the surface;

[0049](5) Reacting the polymer microspheres with amino groups on the surface obtained in step (4) with 5 mL phosphoric acid, and then reacting with 100 mM titanium sulfate at 25° C. for 16 hours to prepare a fixed titanium ion affinity chromatography filler;

[0050](6) Fill the sieve...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
particle diameteraaaaaaaaaa
diameteraaaaaaaaaa
thicknessaaaaaaaaaa
Login to View More

Abstract

The invention relates to a preparation method of a centrifuge-assisted phosphopeptide solid-phase extraction small column. By centrifugation-assisted method, the synthesized immobilized titanium ion affinity chromatography filler was filled into a pipette tip with a sieve plate near the suction port to prepare a phosphopeptide solid-phase extraction column, and the column was used for Enrichment of phosphopeptides in digests of proteomic samples. The preparation method of the phosphopeptide solid-phase extraction small column is simple, low in cost, easy to operate, and has high specificity for the enrichment of the phosphopeptide.

Description

Technical field[0001]The invention belongs to the technical field of a pretreatment method for complex biological samples in the direction of proteomics research, and specifically relates to a preparation method and application of a phosphopeptide solid phase extraction cartridge.Background technique[0002]Protein phosphorylation modification refers to the process of transferring phosphate groups on ATP to protein amino acid residues catalyzed by protein kinases. It is one of the most important covalent modification methods in organisms and the most thorough translation currently studied. After modification. The reversible process of protein phosphorylation and dephosphorylation regulates life activities including cell proliferation, cell differentiation, signal transduction, and cell homeostasis (Reference 1. Yi, T.; Zhai, B. et al. Quantitativephosphoproteomic analysis reveals system-wide signaling pathways downstream of SDF-1 / CXCR4 in breast cancer stem cells, PNAS, 111, E2182-219...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/30B01J20/285B01D15/08G01N30/06
CPCB01D15/08B01J20/285B01J2220/44B01J2220/4806B01J2220/4812G01N30/06
Inventor 叶明亮姚亚婷董靖
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com