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Chromatography of microbial cells using continuous supermacroporous affinity and ion-exchange columns
被引:208
作者:
Arvidsson, P
Plieva, FM
Savina, IN
Lozinsky, VI
Fexby, S
Bülow, L
Galaev, IY
Mattiasson, B
机构:
[1] Lund Univ, Ctr Chem & Chem Engn, Dept Biotechnol, S-22100 Lund, Sweden
[2] Russian Acad Sci, AN Nesmeyanov Organoelement Cpds Inst, Moscow 119991, Russia
[3] Lund Univ, Ctr Chem & Chem Engn, Dept Pure & Appl Biochem, S-22100 Lund, Sweden
[4] Protista Int AB, S-26722 Bjuv, Sweden
关键词:
cells;
stationary phases;
LC;
ion-exchange chromatography;
immobilized metal affinity chromatography;
bacteria;
proteins;
D O I:
10.1016/S0021-9673(02)01114-7
中图分类号:
Q5 [生物化学];
学科分类号:
071010 ;
081704 ;
摘要:
Continuous supermacroporous chromatographic columns with anion-exchange ligands [2-(dimethylanlino)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 mum 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. (C) 2002 Published by Elsevier Science B.V.
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页码:27 / 38
页数:12
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