Fast and efficient protein purification using membrane adsorber systems

verfasst von: Kirstin Suck, Johanna Walter, Frauke Menzel, Alexander Tappe, Cornelia Kasper, Claudia Naumann, Robert Zeidler, Thomas Scheper
Abstract: The purification of proteins from complex cell culture samples is an essential step in proteomic research. Traditional chromatographic methods often require several steps resulting in time consuming and costly procedures. In contrast, protein purification via membrane adsorbers offers the advantage of fast and gentle but still effective isolation. In this work, we present a new method for purification of proteins from crude cell extracts via membrane adsorber based devices. This isolation procedure utilises the membranes favourable pore structure allowing high flow rates without causing high back pressure. Therefore, shear stress to fragile structures is avoided. In addition, mass transfer takes place through convection rather than diffusion, thus allowing very rapid separation processes. Based on this membrane adsorber technology the separation of two model proteins, human serum albumin (HSA) and immungluboline G (IgG) is shown. The isolation of human growth hormone (hGH) from chinese hamster ovary (CHO) cell culture supernatant was performed using a cation exchange membrane. The isolation of the enzyme penicillin acylase from the crude Escherichia coli supernatant was achieved using an anion exchange spin column within one step at a considerable purity. In summary, the membrane adsorber devices have proven to be suitable tools for the purification of proteins from different complex cell culture samples.
Organisationseinheit(en): Institut für Technische Chemie
Externe Organisation(en): Vivascience GmbH
Typ: Artikel
Journal: Journal of biotechnology
Band: 121
Seiten: 361-367
Anzahl der Seiten: 7
ISSN: 0168-1656
Publikationsdatum: 12.09.2005
Publikationsstatus: Veröffentlicht
Peer-reviewed: Ja
ASJC Scopus Sachgebiete: Biotechnologie, Bioengineering, Angewandte Mikrobiologie und Biotechnologie
Elektronische Version(en): https://doi.org/10.1016/j.jbiotec.2005.07.023 (Zugang: Unbekannt)

BibTeX

@article{b8312c58fc2241d69f6158e60423282e,
title = "Fast and efficient protein purification using membrane adsorber systems",
abstract = "The purification of proteins from complex cell culture samples is an essential step in proteomic research. Traditional chromatographic methods often require several steps resulting in time consuming and costly procedures. In contrast, protein purification via membrane adsorbers offers the advantage of fast and gentle but still effective isolation. In this work, we present a new method for purification of proteins from crude cell extracts via membrane adsorber based devices. This isolation procedure utilises the membranes favourable pore structure allowing high flow rates without causing high back pressure. Therefore, shear stress to fragile structures is avoided. In addition, mass transfer takes place through convection rather than diffusion, thus allowing very rapid separation processes. Based on this membrane adsorber technology the separation of two model proteins, human serum albumin (HSA) and immungluboline G (IgG) is shown. The isolation of human growth hormone (hGH) from chinese hamster ovary (CHO) cell culture supernatant was performed using a cation exchange membrane. The isolation of the enzyme penicillin acylase from the crude Escherichia coli supernatant was achieved using an anion exchange spin column within one step at a considerable purity. In summary, the membrane adsorber devices have proven to be suitable tools for the purification of proteins from different complex cell culture samples.",
keywords = "Human growth hormone, Membrane adsorber, Penicillin acylase, Protein purification",
author = "Kirstin Suck and Johanna Walter and Frauke Menzel and Alexander Tappe and Cornelia Kasper and Claudia Naumann and Robert Zeidler and Thomas Scheper",
year = "2005",
month = sep,
day = "12",
doi = "10.1016/j.jbiotec.2005.07.023",
language = "English",
volume = "121",
pages = "361--367",
journal = "Journal of biotechnology",
issn = "0168-1656",
publisher = "Elsevier BV",
number = "3",
}