Abstract
High-resolution protein separation techniques critically depend on the availability of column packings of small average particle size. This gives a minimum of peak broadening on the column owing to the direct relationship between the theoretical plate height parameter, H, and particle size (lowest values of H give highest resolution). High-performance liquid chromatography (HPLC) procedures exploit column packings with average diameters of as little as 5–40 µm. However, these are used in high-pressure systems (up to 400 bar) often with organic solvents and are generally limited to rather low sample loadings (1). To provide a more biocompatible high-resolution separation of biopolymers, including (although not exclusive to) proteins, Pharmacia LKB (Uppsala, Sweden) developed fast protein liquid chromatography (FPLC) in 1982 (2).
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© 1996 Humana Press Inc.
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Sheehan, D. (1996). Fast Protein Liquid Chromatography (FPLC) Methods. In: Doonan, S. (eds) Protein Purification Protocols. Methods in Molecular Biology™, vol 59. Humana Press. https://doi.org/10.1385/0-89603-336-8:269
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DOI: https://doi.org/10.1385/0-89603-336-8:269
Publisher Name: Humana Press
Print ISBN: 978-0-89603-336-8
Online ISBN: 978-1-59259-545-7
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