Fast and efficient protein purification using membrane adsorber systems.
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.
Suck
Kirstin
K
Institut für Technische Chemie der Universität Hannover, Callinstrasse 3, D-30167 Hannover, Germany.
Walter
Johanna
J
Menzel
Frauke
F
Tappe
Alexander
A
Kasper
Cornelia
C
Naumann
Claudia
C
Zeidler
Robert
R
Scheper
Thomas
T
D000327:Adsorption; D000818:Animals; D016466:CHO Cells; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006224:Cricetinae; D017077:Culture Media, Conditioned / Q000737:chemistry; D004586:Electrophoresis; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000201:enzymology; D013006:Growth Hormone / Q000302:isolation & purification*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008567:Membranes, Artificial; D008970:Molecular Weight; D010399:Penicillin Amidase / Q000032:analysis / Q000302:isolation & purification*; D011506:Proteins / Q000302:isolation & purification*; D012709:Serum Albumin / Q000737:chemistry / Q000302:isolation & purification*
D017077:Culture Media, Conditioned; D008567:Membranes, Artificial; D011506:Proteins; D012709:Serum Albumin; D013006:Growth Hormone; D010399:Penicillin Amidase