Abstract Countercurrent chromatographic performance of the locular multilayer coil separation column newly designed in our laboratory, was evaluated in terms of theoretical plate number, peak resolution, and retention of the stationary phase in protein separation with an aqueous polymer phase system, using the small-scale cross-axis coil planet centrifuge (X-axis CPC) fabricated in our laboratory. The locular column was made from 1.0 mm I.D., 2.0 mm O.D., or 1.5 mm I.D., 2.5 mm O.D. PTFE tubing, compressed with a pair of hemostat at 2 or 4 cm intervals. The protein separation was performed using a set of stable proteins including cytochrome C, myoglobin, and lysozyme with the 12.5% (w/w) polyethylene glycol 1000 and 12.5% (w/w) dibasic potassium phosphate system under 1000 rpm of column revolution. The 1.5 mm I.D., 2.5 mm O.D. locular tubing compressed at 2 cm intervals yielded better partition efficiencies than the non-clamped tubing, using both lower and upper mobile phases with satisfactory retention of the stationary phase. The overall results suggest that the newly designed locular multilayer coil is useful to the preparative separation of proteins with aqueous-aqueous polymer phase system using our small scale X-axis CPC.

中文翻译：

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使用具有水-水聚合物相系统的小型交叉轴螺旋行星离心机对蛋白质进行逆流色谱分离的新设计的室性多层螺旋的分配效率

摘要 从理论塔板数、峰分辨率和在聚合物水相系统中分离蛋白质的固定相保留率等方面评价了本实验室新设计的室状多层线圈分离柱的逆流色谱性能，使用小规模我们实验室制造的交叉轴螺旋行星离心机（X轴CPC）。腔镜柱由内径 1.0 毫米、外径 2.0 毫米或内径 1.5 毫米、外径 2.5 毫米的 PTFE 管制成，并用一对止血钳以 2 或 4 厘米的间隔压缩。使用一组稳定的蛋白质（包括细胞色素 C、肌红蛋白和溶菌酶）与 12.5% (w/w) 聚乙二醇 1000 和 12.5% (w/w) 磷酸氢钾系统在 1000 rpm 的色谱柱转速下进行蛋白质分离。1.5 毫米内径，2.5 毫米外径 以 2 cm 间隔压缩的室管比非夹紧管产生更好的分配效率，使用下层和上层流动相，固定相的保留令人满意。总体结果表明，新设计的室状多层线圈可用于使用我们的小规模 X 轴 CPC 对具有水-水聚合物相系统的蛋白质进行制备分离。