Two-dimensional separation for proteomic analysis

Myeong Hee Moon, Ki Hun Kim, Dukjin Kang

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

This chapter describes the development of the two-dimensional separation methods using flow field-flow fractionation (F4) and isoelectric focusing for proteomics utility. The methods described here are the rapid, non-gel-based, on-line, two-dimensional separation methods in which proteins are separated by isoelectric focusing (IEF) in the first dimension according to differences in isoelectric point (pI) followed by size based separation using F4 (either hollow fiber F4 or multilane asymmetrical F4 channels) in an orthogonal direction. In this chapter, the capillary IEF-HF5 and IEF-AF4 methods are described with the demonstration of system performances using protein standards. Also described are the applications to human urinary proteome samples in which proteome fractions are collected and tryptic digested for the proteomic analysis using nanoflow liquid chromatography-tandem mass spectrometry (nLC-ESI-MS-MS).

Original languageEnglish
Title of host publicationField-Flow Fractionation in Biopolymer Analysis
PublisherSpringer-Verlag Wien
Pages57-72
Number of pages16
ISBN (Electronic)9783709101544
ISBN (Print)9783709101537
DOIs
Publication statusPublished - 2012 Jan 1

All Science Journal Classification (ASJC) codes

  • Chemistry(all)

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    Moon, M. H., Kim, K. H., & Kang, D. (2012). Two-dimensional separation for proteomic analysis. In Field-Flow Fractionation in Biopolymer Analysis (pp. 57-72). Springer-Verlag Wien. https://doi.org/10.1007/978-3-7091-0154-4_4