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

Arvidsson, Pär LU ; Plieva, Fatima LU ; Savina, Irina LU ; Lozinsky, VI; Fexby, Sara LU ; Bülow, Leif LU ; Galaev, Igor LU and Mattiasson, Bo LU (2002) In Journal of Chromatography A 977(1). p.27-38
Abstract
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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
metal affinity chromatography, immobilized, ion-exchange chromatography, cells, LC, stationary phases, bacteria, proteins
in
Journal of Chromatography A
volume
977
issue
1
pages
27 - 38
publisher
Elsevier
external identifiers
  • pmid:12456093
  • wos:000179186100004
  • scopus:0037112271
ISSN
0021-9673
DOI
10.1016/S0021-9673(02)01114-7
language
English
LU publication?
yes
id
d714c6d3-8ff7-4a90-8042-b36af34276f4 (old id 324222)
date added to LUP
2007-08-07 15:58:55
date last changed
2017-11-19 04:15:55
@article{d714c6d3-8ff7-4a90-8042-b36af34276f4,
  abstract     = {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.},
  author       = {Arvidsson, Pär and Plieva, Fatima and Savina, Irina and Lozinsky, VI and Fexby, Sara and Bülow, Leif and Galaev, Igor and Mattiasson, Bo},
  issn         = {0021-9673},
  keyword      = {metal affinity chromatography,immobilized,ion-exchange chromatography,cells,LC,stationary phases,bacteria,proteins},
  language     = {eng},
  number       = {1},
  pages        = {27--38},
  publisher    = {Elsevier},
  series       = {Journal of Chromatography A},
  title        = {Chromatography of microbial cells using continuous supermacroporous affinity and ion-exchange columns},
  url          = {http://dx.doi.org/10.1016/S0021-9673(02)01114-7},
  volume       = {977},
  year         = {2002},
}