Chromatography of microbial cells using continuous supermacroporous affinity and ion-exchange columns
Chromatography of microbial cells using continuous supermacroporous affinity and ion-exchange columns
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DOI：
10.1016/s0021-9673(02)01114-7
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发表时间：
2002-11-15
影响因子：
4.1
通讯作者：
Mattiasson, B
中科院分区：
文献类型：
--
作者：
Arvidsson, P;Plieva, FM;Mattiasson, B
连续超大孔色谱柱具有阴离子交换配体[2-(二甲基氨基)乙基]和固定化金属亲和(IMA)配体(负载Cu2+的亚氨基二乙酸)，可与大肠杆菌细胞结合并以高回收率洗脱结合细胞。这些基于聚丙烯酰胺的连续超大孔柱是通过单体在冻结在柱内的水溶液中自由基共聚合(冷冻聚合)而得到的。解冻后，柱子包含一个连续的基质(所谓的冷冻胶)，具有大小为10-100微米的相互连接的孔。基质的大孔径使大肠杆菌细胞能够畅通无阻地通过不含配体的平坦柱子。分别用10 mM咪唑或20 mM EDTA溶液洗脱低离子强度离子交换柱上的大肠杆菌细胞，在0.35-0.40M的氯化钠浓度下洗脱回收率为70-80%，而洗脱IMA柱上的细胞，回收率为80%左右。细胞在结合/洗脱过程后保持其活性。这些初步结果表明，微生物细胞可以在使用超大孔连续柱的色谱模式下处理。这些色谱柱很容易用廉价且容易获得的原料制造，这使得这些色谱柱适合一次性使用。(C)2002年，爱思唯尔科学公司出版。
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.