Chromatography of microbial cells using continuous supermacroporous affinity and ion-exchange columns
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/324222
- author
- Arvidsson, Pär LU ; Plieva, Fatima LU ; Savina, Irina LU ; Lozinsky, VI ; Fexby, Sara LU ; Bülow, Leif LU ; Galaev, Igor LU and Mattiasson, Bo LU
- organization
- publishing date
- 2002
- 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
- 2016-04-01 17:14:48
- date last changed
- 2022-04-15 18:15:43
@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}}, keywords = {{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}}, doi = {{10.1016/S0021-9673(02)01114-7}}, volume = {{977}}, year = {{2002}}, }