In-situ rapid preprocessing method for protein samples

A protein and pretreatment technology, applied in the preparation of test samples, etc., can solve the problems of large loss, cumbersome operation, and time-consuming, and achieve the effect of reducing loss, reducing desalination steps, and reducing sample loss.

Active Publication Date: 2014-10-01
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
View PDF5 Cites 25 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to overcome the shortcomings of the protein sample pretreatment process, such as time-consuming, cumbersome operation, and large loss, the present invention provides a protein sample in-situ rapid pretreatment method, which integrates protein denaturation, reduction, solvent replacement, Alkylation, enzymatic hydrolysis and impurity removal for the entire sample pretreatment process

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
  • In-situ rapid preprocessing method for protein samples
  • In-situ rapid preprocessing method for protein samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0023] The centrifugal ultrafiltration device that the present invention adopts is commercialization (purchased from Millipore, Billerica, MA) and is made up of ultrafiltration tube and collection tube; Cylindrical structure of regenerated cellulose membrane);

[0024] When in use, the ultrafiltration tube is placed in the collection tube.

[0025] 1. Preparation of Protein Solution

[0026] Escherichia coli protein samples (purchased from Sigma, St.Louis, MO) were dissolved in 10mM ammonium bicarbonate buffer solution (ABC) (pH8.7), and after the concentration was determined by BCA method, the protein sample concentration was prepared with 10mM ABC as solvent 1 mg / mL.

[0027] 2. Denaturation and reduction of protein solutions

[0028] Take 30 μL of E. coli protein solution prepared in step 1 (i.e. 30 μg sample) and place it in an ultrafiltration tube, add 10 mM ABC to make up to 60 μL solution, add 1M dithiothreitol (DTT) (dissolved in 10 mMABC) 6 μL ( The final concent...

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

No PUM Login to view more

Abstract

The invention relates to an in-situ rapid preprocessing method for protein samples. The whole preprocessing process of the protein samples is carried out in a centrifugal ultrafiltration device in situ, the denaturation process, the reduction process, the solvent replacement process, the alkylate process, the enzymolysis process and the impurity removal process of the protein samples are integrated, and enzymolysis of the protein samples is accelerated under assistance of microwaves. The method can remarkably improve the preprocessing flux of the protein samples; meanwhile, the whole preprocessing process of the samples is carried out in situ, and the recovery rate is high. In addition, the centrifugal ultrafiltration device can conveniently and rapidly achieve the purification of the samples and replacement of solvents, and enzymatic hydrolysate does not need an additional purification process and can be directly combined with subsequent efficient liquid chromatogram separation and mass spectrum identification.

Description

technical field [0001] The invention relates to a rapid in-situ pretreatment method for protein samples. Background technique [0002] The identification of proteins is closely related to the health of the social population. For most genetic diseases, such as tumors and obesity, certain proteins are differentially expressed between patients and healthy people. In order to achieve "early detection and early treatment" of diseases, the rapid and highly sensitive identification of specific proteins is of great significance. As the first step in protein analysis, protein sample pretreatment can achieve the removal of impurities and the enrichment of samples. Therefore, the establishment of an efficient and fast protein sample pretreatment method will play a role in the research process of achieving high-sensitivity and high-accuracy differential proteins. It plays an important role, and then promotes the in-depth research on the discovery and treatment of diseases. [0003] C...

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 Applications(China)
IPC IPC(8): G01N1/44G01N1/34
Inventor 张丽华方菲赵群杨开广张玉奎
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap