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.
引用
收藏
页码:27 / 38
页数:12
相关论文
共 22 条
[1]   FLOW-THROUGH PARTICLES FOR THE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC SEPARATION OF BIOMOLECULES - PERFUSION CHROMATOGRAPHY [J].
AFEYAN, NB ;
GORDON, NF ;
MAZSAROFF, I ;
VARADY, L ;
FULTON, SP ;
YANG, YB ;
REGNIER, FE .
JOURNAL OF CHROMATOGRAPHY, 1990, 519 (01) :1-29
[2]  
Albertson P. A., 1986, PARTITION CELL PARTI
[3]  
ARVIDSSON P, 2002, UNPUB J CHROMATOGR A
[4]  
Braas G, 1996, BIOSEPARATION, V6, P211
[5]   Recovery and manipulation of nanoparticulate bioproducts: Relevance to the up-scaled manufacture of gene therapy vectors [J].
Braas, GMF ;
Walker, SG ;
Zhang, Z ;
Lyddiatt, A .
FOOD AND BIOPRODUCTS PROCESSING, 2000, 78 (C1) :11-18
[6]   PURIFICATION OF PROTEINS BY ADSORPTION CHROMATOGRAPHY IN EXPANDED BEDS [J].
CHASE, HA .
TRENDS IN BIOTECHNOLOGY, 1994, 12 (08) :296-303
[7]   Direct measurements of convective fluid velocities in superporous agarose beads [J].
Gustavsson, PE ;
Axelsson, A ;
Larsson, PO .
JOURNAL OF CHROMATOGRAPHY A, 1998, 795 (02) :199-210
[8]   Continuous superporous agarose beds for chromatography and electrophoresis [J].
Gustavsson, PE ;
Larsson, PO .
JOURNAL OF CHROMATOGRAPHY A, 1999, 832 (1-2) :29-39
[9]   Peak broadening in protein chromatography with monoliths at very fast separations [J].
Hahn, R ;
Jungbauer, A .
ANALYTICAL CHEMISTRY, 2000, 72 (20) :4853-4858
[10]   Fast isolation of protein receptors from streptococci G by means of macroporous affinity discs [J].
Kasper, C ;
Meringova, L ;
Freitag, R ;
Tennikova, T .
JOURNAL OF CHROMATOGRAPHY A, 1998, 798 (1-2) :65-72