title: Chromatography of microbial cells using continuous supermacroporous affinity and ion-exchange columns.
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 microm 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.
authors.affiliation: Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, S-22100 Lund, Sweden.
authors.forename: Pär
authors.initials: P
authors.lastname: Arvidsson
authors.forename: Fatima M
authors.initials: FM
authors.lastname: Plieva
authors.forename: Irina N
authors.initials: IN
authors.lastname: Savina
authors.forename: Vladimir I
authors.initials: VI
authors.lastname: Lozinsky
authors.forename: Sara
authors.initials: S
authors.lastname: Fexby
authors.forename: Leif
authors.initials: L
authors.lastname: Bülow
authors.forename: Igor Yu
authors.initials: IY
authors.lastname: Galaev
authors.forename: Bo
authors.initials: B
authors.lastname: Mattiasson
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004926:Escherichia coli / Q000302:isolation & purification*; D008855:Microscopy, Electron, Scanning