Spiral counter-current chromatography of small molecules, peptides and proteins using the spiral tubing support rotor.
Spiral counter-current chromatography of small molecules, peptides and proteins using the spiral tubing support rotor.
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DOI：
10.1016/j.chroma.2011.06.007
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发表时间：
2011-09-09
期刊：
影响因子：
--
通讯作者：
Pilon A
中科院分区：
文献类型：
--
作者：
Knight M;Finn TM;Zehmer J;Clayton A;Pilon A
逆流色谱（CCC）的一个重要进展是在行星式离心机中旋转的开流管盘管中进行的，这是一种螺旋展开管的新设计。发现管之间的更大间距显著增加固定相保留，使得现在所有类型的两相溶剂系统都可以用于J型行星式离心机中的液-液分配色谱。通过激光烧结技术构建具有圆形通道的螺旋管支撑（STS）框架，其中FEP管从底部到顶部以每层4个螺旋环放置，并且附连盖，允许管连接到行星式离心机的流管。转子安装并在PC中运行。Inc.型仪器。在适当的两相溶剂系统中对分子量在<300至约15，000范围内的化合物的实例进行色谱分析，以评估分离和纯化的能力。在正己烷-乙酸乙酯-甲醇-水介质中，阿司匹林等小分子物质得到了完全分离。在仲丁醇溶剂系统中将包括非常疏水的肽的合成肽各自纯化至非常高的纯度水平。在STS转子高固定相保留是可能的，在正常的流速与含水仲丁醇溶剂系统。最后，将聚乙二醇-磷酸钾两相水溶液体系用于分离大肠杆菌裂解液中的蛋白质。coli表达系统。这些实验证明了使用STS转子的螺旋CCC的多功能性。
An important advance in countercurrent chromatography (CCC) carried out in open flow-tubing coils, rotated in planetary centrifuges, is the new design to spread out the tubing in spirals. More spacing between the tubing was found to significantly increase the stationary phase retention, such that now all types of two-phase solvent systems can be used for liquid-liquid partition chromatography in the J-type planetary centrifuges. A spiral tubing support (STS) frame with circular channels was constructed by laser sintering technology into which FEP tubing was placed in 4 spiral loops per layer from the bottom to the top and a cover affixed allowing the tubing to connect to flow-tubing of the planetary centrifuge. The rotor was mounted and run in a P.C. Inc. type instrument. Examples of compounds of molecular weights ranging from <300 to approximately 15,000 were chromatographed in appropriate two-phase solvent systems to assess the capability for separation and purification. A mixture of small molecules including aspirin was completely separated in hexane-ethyl acetate-methanol-water. Synthetic peptides including a very hydrophobic peptide were each purified to a very high purity level in a sec-butanol solvent system. In the STS rotor high stationary phase retention was possible with the aqueous sec-butanol solvent system at a normal flow rate. Finally, the two-phase aqueous polyethylene glycol-potassium phosphate solvent system was applied to separate a protein from a lysate of an E. coli expression system. These experiments demonstrate the versatility of spiral CCC using the STS rotor.
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DOI：
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发表时间：
2009-05-08
期刊：
Journal of chromatography. A
影响因子：
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作者：
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影响因子：
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作者：
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发表时间：
2006-12-05
影响因子：
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作者：
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