Institute for Neuroanatomy, Centre for Biomedicine and Medical Technology Mannheim, Medical Faculty Mannheim, University of Heidelberg, Heidelberg, Germany.
FFE Service GmbH, Feldkirchen, Germany.
Electrophoresis. 2018 Sep;39(18):2288-2299. doi: 10.1002/elps.201800187. Epub 2018 Jun 6.
Free-flow electrophoresis (FFE) exploits differences in the overall charge of bio-particles separating cells, organelles, macromolecules, ions, etc. according to their distinct electrophoretic mobility and isoelectric point (pI) values. Indeed, around a neutral pH organelles usually exhibit a negative surface charge, migrating in an electric field from the cathode toward the anode. Since its introduction more than five decades ago by Barrollier et al., Z. Naturforsch. 1958, 13b, 745-755 and Hannig, Z. Anal. Chem. 1961, 181, 244-254, FFE has become an established analytical and preparative separation method for the isolation of a variety of organelles. Particularly, in sophisticated, multistep separating processes to separate subpopulations of organelles, it has gained, meanwhile, a position as a versatile technology and essential element. Relying on the distinct surface charges instead of buoyant densities of cell organelles, the FFE technology is best supporting a preceding centrifugation-based fractionation of subcellular compartments in the second dimension. In the following review, the two-step isolation and purification of subpopulations of classic animal and plant cell organelles will be mainly exemplified.
自由流电泳（FFE）利用生物颗粒的总电荷差异，根据其独特的电泳迁移率和等电点（pI）值来分离细胞、细胞器、大分子、离子等。事实上，在中性 pH 左右，细胞器通常带有负表面电荷，在电场中从阴极向阳极迁移。自五十多年前由 Barrollier 等人首次引入以来，Z. Naturforsch. 1958, 13b, 745-755 和 Hannig, Z. Anal. Chem. 1961, 181, 244-254，FFE 已成为一种成熟的分析和制备分离方法，用于分离各种细胞器。特别是在复杂的多步分离过程中，用于分离细胞器的亚群，它已经成为一种通用技术和必不可少的元素。FFE 技术依赖于细胞器的表面电荷而不是浮力密度，最适合在二维空间中对亚细胞区室进行基于离心的分级分离。在接下来的综述中，将主要举例说明经典动植物细胞细胞器亚群的两步分离和纯化。
