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

A dual-channel spe column and its application in quantitative proteomics

A dual-channel, protein technology, applied in measurement devices, scientific instruments, instruments, etc., can solve the problems of low sample recovery rate, sample loss and contamination, and inability to respond to chemically labeled samples, etc. Effect of sample loss

Active Publication Date: 2016-01-27
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The sample pretreatment process of this method includes protein extraction, urea denaturation, dithiothreitol (DTT) reduction, iodoacetamide (IAA) alkylation, trypsin digestion and dimethylation labeling, stop labeling and C18 reaction. Compared with desalting, so many operations are prone to sample loss and contamination
Existing solution is filter-assisted sample preparation (FASP) (NatMethods, 2009.6: 359), but this method sample recovery rate is low and can't be applied to the analysis of chemically labeled sample; Heck etc. realize dimethylation mark in SPE ( Nature Protocols, 2009.4:484), but this method ignores the loss of samples before labeling

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 dual-channel spe column and its application in quantitative proteomics
  • A dual-channel spe column and its application in quantitative proteomics
  • A dual-channel spe column and its application in quantitative proteomics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0032] In order to investigate the effect of dual-channel SPE on protein quantification, the device was used to process protein samples. The specific steps are as follows:

[0033] 1. The production process of dual-channel SPE: divide the 1mL SPE tube into two equal parts with a paper cutter, add a sieve plate at the bottom of the channel, use a polytetrafluoroethylene film as a separator in the middle of the channel, and use a plastic glue stick to separate the separator. The plate is sealed with the SPE column tube, and filled with C18 reversed-phase chromatography filler (40 μm particle size).

[0034] 2. Protein sample processing process: Add 100 μL of 0.1 μg / μL BSA dissolved in 8M urea to each of the two channels, and add 1 μL of 0.1mol / L dithiothreitol to react at room temperature for 1 hour, then add 2 μL of 0.1M iodoethyl The amide was reacted in the dark for 30 minutes, and then diluted 10 times with 50mM phosphate buffer (pH8.0), then added trypsin according to 1:20 ...

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
diameteraaaaaaaaaa
diameteraaaaaaaaaa
particle diameteraaaaaaaaaa
Login to View More

Abstract

The invention relates to a novel solid phase extraction (SPE) device capable of achieving the whole process of quantitative analysis pretreatment of protein. According to the device, an SPE tube is partitioned into two chambers with the same capacity through a partition plate, and after the chambers are filled with chromatographic packing, the whole process of sample treatment in quantitative proteomics can be achieved in situ, wherein the treatment comprises denaturation, reduction, alkylation, enzymolysis, isotope labeling and mixing of protein samples. The device is easy and convenient to manufacture and low in manufacturing cost, and has the advantages that the recovery rate is high, operation is easy, the accuracy of quantitative results is good and the precision is high when the samples are treated.

Description

technical field [0001] The present invention designs a dual-channel SPE device, which can realize the whole process of quantitative protein sample processing such as protein sample in-situ denaturation, reduction, alkylation, enzymatic hydrolysis, labeling, desalting and mixing. Background technique [0002] Quantitative proteomics is a hot topic in systems biology research. It provides extremely important data for discovering disease biomarkers, discovering protein-protein interactions, drug-protein interactions, explaining the mechanism of disease, explaining biological metabolic processes, etc. support. Protein is the embodiment and bearer of life activities. It is extremely rich in organisms and participates in all metabolic processes of living organisms. Therefore, changes in protein content will lead to physiological changes, and for humans, it is reflected in the occurrence of diseases. Therefore Accurate quantification of proteins is extremely important for understa...

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): G01N30/60
Inventor 张丽华周愿赵群张珅杨开广张玉奎
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