Novel analytical spiral tube assembly for protein separation by high-speed countercurrent chromatography

1.介绍了一种用于高速逆流色谱分析分离的新型螺旋柱组件。柱子由塑料螺旋管支架制成，螺旋管支架上有12个交织的螺旋槽。首先将直径为1.6 mm的聚四氟乙烯管材通过狭缝挤出变平，然后插入凹槽中，形成5层螺旋层，容量约为60ml。在聚乙二醇1000和磷酸二氢钾组成的聚合物相体系中分离了三种稳定的蛋白质样品，包括细胞色素C、肌红蛋白和溶菌酶，测试了螺旋柱组件的性能，每个样品在水中的质量分数为12.5%。在2ml/min的速度下，3个蛋白质样品在1小时内得到很好的分解。今后在提高螺旋管支架强度后，采用较高的转速和流量，可进一步缩短分离时间。 2.新型螺旋管组件用于蛋白质的高速逆流半制备分离 描述了一种用于蛋白质高速逆流色谱制备分离的新型螺旋柱组件。柱子由塑料螺旋管支架制成，该支架有4个交织的螺旋槽和12个径向沟槽，以容纳相互连接的管子。两个分离柱是虚构的。一种是将直径1.6 mm的聚四氟乙烯管插入螺旋槽中，形成13层螺旋层，容量为135ml。另一种螺旋柱是由同一管子制成的，首先将其挤出一条狭缝，然后沿其轴线扭曲形成平扭管，将其插入螺旋槽形成10个螺旋层，总容量为90毫升。这些螺旋柱组件首先在由聚乙二醇1000和二盐酸钾组成的聚合物相体系中分离细胞色素C、肌红蛋白和溶菌酶，在水中以1、2、3和5ml/min的速度旋转，转速分别为800和1000rpm。三个蛋白质样品在1000rpm时在两个色谱柱中都得到了很好的分离，而改进的平扭曲色谱柱的分离效率略高。在进一步提高螺旋管支架强度后，采用更高的转速和流量，可以进一步提高分离效果。

1. A new spiral column assembly for analytical separation by high-speed countercurrent chromatography is described. The column is made from a plastic spiral tube support which has 12 interwoven spiral grooves. The PTFE tubing of 1.6 mm ID was first flattened by extruding through a narrow slit and inserted into the grooves to make 5 spiral layers with about 60 ml capacity. The performance of the spiral column assembly was tested with the separation of three stable protein samples including cytochrome C, myoglobin and lysozyme in a polymer phase system composed of polyethylene glycol 1000 and dibasic potassium phosphate each at 12.5 % (w/w) in water. At 2 ml/min, three protein samples were well resolved in one hour. The separation tme may be further shortened by application of higher revolution speed and flow rate after improving the strength of the spiral tube support in the future. 2. Novel spiral tube assembly for semipreparative sparation of proteins by high-speed countercurrent chromatography A new spiral column assembly for preparative separation of proteins by high-speed countercurrent chromatography is described. The column is made from a plastic spiral tube support which has 4 interwoven spiral grooves with 12 radial grooved to accommodate interconnecting tubing. Two separation columns was fabicated. One was made from PTFE tubing of 1.6 mm ID by inserting it into the spiral grooves to form 13 spiral layers with 135 ml capacity. The other column was made from the same tubing by first flattening by extruding it through a narrow slit followed by twisting along its axis to form flat-twisting tubing by inserting it into spiral grooves to form 10 sprial layer with a total capacity of 90 ml. The performance of these spiral column assemblies were first tested on the separation of three stable protein samples including cytochrome C, myoglobin and lysozyme in a polymer phase system composed of polyethylene glycol 1000 and dibasic potassium phosphate each at 12.5 % (w/w) in water at flow rates of 1, 2, 3 and 5 ml/min and the revolution speeds of 800 and 1000 rpm. Three protein samples were well resolved in both columns at 1000 rpm, while the modified flat-twisted column yielded slightly higher separaton efficiency. The separation may be further improved by application of higher revolution speed and flow rate after improving the strength of the spiral tube support in the future.