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Counterbalancing Hydrodynamic Sample Distortion Effects Increases Resolution of Free‐flow Zone Electrophoresis by Dr. Gerhard Weber; Johann Bauer is a Engineering article available to read on EtoBox.

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## Abstract On fractionation of highly heterogeneous protein mixtures, optimal resolution was achieved by forcing proteins to migrate through a preestablished pH gradient, until they entered a medium with a pH similar but not equal to their p__I__s. For this purpose, up to seven different media were pumped through the electrophoresis chamber so that they were flowing adjacently to each other, forming a pH gradient declining stepwise from the cathode to the anode. This gradient had a sufficiently strong band‐focusing effect to counterbalance sample distortion effects of the flowing medium as proteins approached their isoelectric medium closer than 0.5 pH units. Continuous free‐flow zone electrophoresis (FFZE) with high throughput capability was applicable if proteins did not precipitate or aggregate in these media. If components of heterogeneous protein mixtures had already started to precipitate or aggregate, in a medium with a pH exceeding their p__I__ by more than 0.5 pH units, the application of interval modus and media forming flat pH gradients appeared advantageous.

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Author

Dr. Gerhard Weber; Johann Bauer

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Ltd.; Wiley (ISSN 0173-0835)

Published

1998

Language

EN

Field

Engineering (Physical Sciences)