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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Official URL: http://dx.doi.org/10.1016/s0021-9673(02)01114-7
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.
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