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An integrated method for the detection of basic proteins in serum-derived proteomes

Paper ID Volume ID Publish Year Pages File Format Full-Text
34143 45005 2016 8 PDF Available
Title
An integrated method for the detection of basic proteins in serum-derived proteomes
Abstract

•The difficulty of serum proteomics application for the basic regions of proteins.•Integrating approach by using Micro-Rotofor IEF fractionation and AU-PAGE.•Successful detections for C3 and C4A complement fragments proteins.•Basic protein detection in atopic dermatitis patients-derived sera samples.

Because serum is an optimal source to identify proteins derived from diseased-tissue compartments, serum proteomics has been applied for the discovery and analysis of biomarkers related to human disease. The general 2D-PAGE method is suitable for acidic and neutral proteome separation while highly basic proteins remain unresolved. In this study, we optimized basic proteome fractionation by integrating several methods such as, Micro-Rotofor isoelectric focusing (IEF) fractionation, acetic acid-urea polyacrylamide gel electrophoresis (A-U PAGE) separation, and mass spectrometry analysis for identifying basic proteins in patient sera. The sera samples were obtained from patients with atopic dermatitis (AD) to establish the model method introduced in the present study. We successfully identified the specific presence of a C3 complement component fragment protein and C4A fragment protein in AD patient-derived sera. Highly basic proteins in serum are very difficult to detect due to their separation properties. Thus, the integrated proteomic method approaches described here could be applicable for the detection of basic proteins associated with other diseases.

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Keywords
AD, atopic dermatitis; ADe, extrinsic type of AD; 2D-PAGE, two-dimensional polyacrylamide gel electrophoresis; MALDI-TOF, matrix-assisted laser desorption/ionization-time of flight; IPG, immobilized pH gradient; MARC, multiple affinity removal column; IEF
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An integrated method for the detection of basic proteins in serum-derived proteomes
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 51, Issue 8, August 2016, Pages 973–980
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us