Development of non-gel-based two-dimensional separation of intact proteins by an on-line hyphenation of capillary isoelectric focusing and hollow fiber flow field-flow fractionation
Development of non-gel-based two-dimensional separation of intact proteins by an on-line hyphenation of capillary isoelectric focusing and hollow fiber flow field-flow fractionation
Engineering Village 2
2006 Elsevier Inc.
Accession number: 063510086932
Title: Development of non-gel-based two-dimensional separation of intact proteins by an on-line hyphenation of capillary isoelectric focusing and hollow fiber flow field-flow fractionation
Authors: Kang, Dukjin; Moon, Myeong Hee
Author affiliation: Department of Chemistry, Yonsei University, Seoul, 120-749, South Korea
Serial title: Analytical Chemistry
Abbreviated serial title: Anal. Chem.
Volume: v 78
Issue: n 16
Issue date: Aug 15 2006
Publication year: 2006
Pages: p 5789-5798
Language: English
ISSN: 0003-2700
CODEN: ANCHAM
Document type: Journal article (JA)
Publisher: American Chemical Society, Columbus, OH 43210-3337, United States
Abstract: A rapid, non-gel-based, on-line, two-dimensional separation method is introduced for proteome analysis. Protein fractionation was carried out by first exploiting the differences in their respective isoelectric points (pI) in a Teflon capillary using isoelectric focusing (IEF), followed by a molecular weight (MW)-based separation in a hollow fiber by flow field-flow fractionation (F1FFF). The method developed here (CIEF-HFF1FFF) may be a powerful alternative to two-dimensional polyacrylamide gel electrophoresis, which is currently used for the separation and purification of proteins. In CIEF-HFF1FFF, proteins can be collected as a fraction of a certain pI and MW interval without being denatured. Additionally, the ampholyte solution is simultaneously removed during separation in the hollow fiber, and the overall process time is significantly reduced. This method was applied to a human urinary proteome sample, leading to the identification of 114 proteins with the subsequent off-line use of nanoflow liquid chromatography-tandem mass spectrometry after the tryptic digestion of each collected protein fraction. ? 2006 American Chemical Society.
Number of references: 38
Ei main heading: Proteins
Ei controlled terms: Gels - Separation - Synthetic fibers - Fractionation
Uncontrolled terms: Hyphenation - Isoelectric focusing (IEF) - Liquid chromatography-tandem mass spectrometry
Ei classification codes: 804.1 Organic Compounds - 801.3 Colloid Chemistry - 802.3 Chemical Operations - 819.2 Synthetic Fibers - 811.0.3 Economics, Research and Miscellaneous - 819.5.1 Cellulosic Textiles - 811.0.2 Chemistry, Physics and Mathematics
Treatment: Experimental (EXP)
DOI: 10.1021/ac0606958
Database: Compendex
Compilation and indexing terms, ? 2006 Elsevier Inc. All rights reserved
Subscription required: http://www.engineeringvillage2.org
Votes:23
Comments: 0