Chromatography of microbial cells using continuous supermacroporous affinity and ion-exchange columns

Arvidsson, P., Plieva, F.M., Savina, I.N., Lozinsky, V.I, Fexby, S., Bulow, L., Galaev, I.Y. and Mattiasson, B. (2002) Chromatography of microbial cells using continuous supermacroporous affinity and ion-exchange columns Journal of Chromatography A, 977 (1). pp. 27-38. ISSN 0021-9673

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Abstract

Continuous supermacroporous chromatographic columns with anion-exchange ligands [2-(dimethylamino)ethyl group] and immobilized metal affinity (IMA) ligands (Cu2+-loaded iminodiacetic acid) have been developed allowing binding of Escherichia coli cells and the elution of bound cells with high recoveries. These poly(acrylamide)-based continuous supermacroporous columns have been produced by radical co-polymerization of monomers in aqueous solution frozen inside a column (cryo-polymerization). After thawing, the column contains a continuous matrix (so-called cryogel) with interconnected pores of 10-100 m in size. The large pore size of the matrix makes it possible for E. coli cells to pass unhindered through a plain column containing no ligands. E. coli cells bound to an ion-exchange column at low ionic strength were eluted with 70-80% recovery at NaCl concentrations of 0.35-0.40 M, while cells bound to an IMA-column were eluted with around 80% recovery using either 10 mM imidazole or 20 mM EDTA solutions, respectively. The cells maintain their viability after the binding/elution procedure. These preliminary results indicate that microbial cells can be handled in a chromatographic mode using supermacroporous continuous columns. These columns are easy to manufacture from cheap and readily available starting materials, which make the columns suitable for single-time use.

Item Type: Journal article
Subjects: C000 Biological and Biomedical Sciences
DOI (a stable link to the resource): 10.1016/s0021-9673(02)01114-7
Faculties: Faculty of Science and Engineering > School of Pharmacy and Biomolecular Sciences
Depositing User: editor spbs
Date Deposited: 08 Oct 2007
Last Modified: 21 Oct 2014 16:08
URI: http://eprints.brighton.ac.uk/id/eprint/2678

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