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

P. Arvidsson, F.M. Plieva, Irina Savina, V.I Lozinsky, S. Fexby, L. Bulow, I.Y. Galaev, B. Mattiasson

Research output: Contribution to journalArticleResearchpeer-review

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.
Original languageEnglish
Pages (from-to)27-38
Number of pages12
JournalJournal of Chromatography A
Volume977
Issue number1
DOIs
Publication statusPublished - 15 Nov 2002

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Chromatography
Escherichia coli
Ion exchange
Ligands
Recovery
Cryogels
Metals
Thawing
Polyacrylates
Ionic strength
Edetic Acid
Copolymerization
Pore size
Anions
Monomers
Polymerization

Cite this

Arvidsson, P. ; Plieva, F.M. ; Savina, Irina ; Lozinsky, V.I ; Fexby, S. ; Bulow, L. ; Galaev, I.Y. ; Mattiasson, B. / Chromatography of microbial cells using continuous supermacroporous affinity and ion-exchange columns. In: Journal of Chromatography A. 2002 ; Vol. 977, No. 1. pp. 27-38.
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Arvidsson, P, Plieva, FM, Savina, I, Lozinsky, VI, Fexby, S, Bulow, L, Galaev, IY & Mattiasson, B 2002, 'Chromatography of microbial cells using continuous supermacroporous affinity and ion-exchange columns', Journal of Chromatography A, vol. 977, no. 1, pp. 27-38. https://doi.org/10.1016/s0021-9673(02)01114-7

Chromatography of microbial cells using continuous supermacroporous affinity and ion-exchange columns. / Arvidsson, P.; Plieva, F.M.; Savina, Irina; Lozinsky, V.I; Fexby, S.; Bulow, L.; Galaev, I.Y.; Mattiasson, B.

In: Journal of Chromatography A, Vol. 977, No. 1, 15.11.2002, p. 27-38.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Arvidsson, P.

AU - Plieva, F.M.

AU - Savina, Irina

AU - Lozinsky, V.I

AU - Fexby, S.

AU - Bulow, L.

AU - Galaev, I.Y.

AU - Mattiasson, B.

PY - 2002/11/15

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N2 - 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.

AB - 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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DO - 10.1016/s0021-9673(02)01114-7

M3 - Article

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JO - Journal of Chromatography A

JF - Journal of Chromatography A

SN - 0021-9673

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