REVIEW: New Methods of Protein Purification. Expanded Bed Chromatography
I. Yu. Galaev
Department of Biotechnology, Center for Chemistry and Chemical
Engineering, Lund University, P.O. Box 124, S-22100, Lund, Sweden; fax:
46-46-224713; E-mail:
biot-iga@hermes.lu.se
Received October 29, 1997; Revision received December 12, 1997
This review considers a new method of protein purification--expanded bed
chromatography. The method is based on pumping of the mobile phase
upwards through the column bed. The bed starts to expand at liquid flow
rate above a critical value, and the gaps between the sorbent beads
expand. The controlled distribution of bead size and weight results in
a stable expanded bed, where separate beads are not completely
immobile, but oscillate about some steady position. The mobile phase
flow through such a column is similar to plug flow. The main advantage
of expanded bed chromatography is that it enables protein recovery
directly from particulate-containing feed stocks like cell homogenates
or even fermentation broth, when extracellular proteins are purified.
The successful examples of protein purification by expanded bed
chromatography and the available commercial systems are presented. The
standard equipment for liquid protein chromatography can be used in the
expanded bed mode.
KEY WORDS: protein purification, affinity chromatography,
expanded bed chromatography