Free‐flow electrophoresis in the proteomic era: A technique in flux

Abstract

Since its introduction five decades ago, free-flow electrophoresis (FFE) has been mainly employed for the isolation and fractionation of cells, cell organelles and protein mixtures. In the meantime, the growing interest in the proteome of these bio-particles and biopolymers has shed light on two further facets in the potential of FFE, namely its applicability as an analytical tool and sensor. This review is intended to outline recent innovations, FFE has gained in the proteomic era, and to point out the valuable contributions it has made to the analysis of the proteome of cells, sub-cellular organelles and functional protein networks.

Isolation and Proteomic Characterization of the Arabidopsis Golgi Defines Functional and Novel Components Involved in Plant Cell Wall Biosynthesis

“…The specific protein and lipid constituents of membranes result in variation in surface charge between organelles and therefore different migration distances in an electric field. These characteristics are exploited by free-flow electrophoresis (FFE), in which a mixture of membranes and organelles is introduced into a chamber, moving up under laminar flow while an electric field is applied at right angles to the direction of flow (Islinger et al, 2010). In combination with other organelle-enrichment techniques, FFE has been used successfully in the preparation of a variety of organelles from several plant species, including tonoplast and plasma membrane vesicles at high levels of purity, but to date has not been used in the isolation of Golgi vesicles (Canut et al, 1988(Canut et al, , 1990Bardy et al, 1998).…”

Proteomics and Metabolomics of Marine Organisms: Current Strategies and Knowledge

“…A quite different and uncommon strategy was used for the cyanobacterium Nostoc punctiforme (Wase et al, 2012), based on a free-flow electrophoresis (Islinger, Eckerskorn, and V€ olkl, 2010)-mediated separation of the proteins before digestion.…”

The Current State of the Golgi Proteomes

“…The apparatus can be operated in two modes: zonal electrophoresis (ZE), or isoelectric focusing (IEF) mode. ZE-FFE is becoming recognized for its impressive separation and purification capacity of plant, mammalian and yeast organelles (reviewed by Islinger et al, 2010). The first use of FFE for Golgi was applied to mammalian Golgi membranes and lead to separation of sub-Golgi compartments, demonstrated by a series of enzyme assays (Hartelschenk et al, 1991).…”

“…The first use of FFE for Golgi was applied to mammalian Golgi membranes and lead to separation of sub-Golgi compartments, demonstrated by a series of enzyme assays (Hartelschenk et al, 1991). However, this was prior to the proteomic era and was never The diagram outlines a late commercially model available through BD Diagnostics, with counter flow at sample outlets and stabilization buffers at the extreme anodic and cathodic carrier buffer inlets (Islinger et al, 2010). MicroFFE apparatus are similar with 56.5 mm ×35 mm × 30 mm dimensions (Turgeon & Bowser, 2009).…”