One-step purification of cholinesterase from human serum by CCC
One-step purification of cholinesterase from human serum by CCC
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DOI：
10.1081/jlc-100104375
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发表时间：
2001-01-01
影响因子：
1.3
通讯作者：
Ito, Y
中科院分区：
文献类型：
--
作者：
Shibusawa, Y;Hosojima, T;Ito, Y
XL 型十字轴线圈行星离心机 (CPC) 成功用于通过逆流色谱 (CCC) 从人血清中纯化胆碱酯酶 (ChE)。由聚乙二醇-磷酸钾缓冲液组成的水-水聚合物相系统在各种 pH 值（范围为 6.8 至 9.2）下测试了 ChE、人血清白蛋白、α 球蛋白和 γ 球蛋白的分配比 (D)。 CCC 分离是在两个不同的盘管柱中使用优化的聚合物相系统进行的。使用 16.0% PEG 600-12.5% 磷酸钾在 pH 9.2 下实现最佳纯化，通过小容量柱以 0.5 mL/min 洗脱下层相，其中 ChE 在 2 小时内直接从人血清中纯化。通过 SDS PAGE 分析，酶活性证实的 ChE 级分不含血清蛋白。
The type XL cross-axis coil planet centrifuge (CPC) was successfully utilized for purification of cholinesterase (ChE) from human serum by countercurrent chromatography (CCC). Aqueous-aqueous polymer phase systems composed of polyethylene glycol-potassium phosphate buffers were tested at various pHs, ranging from 6.8 to 9.2 for the distribution ratios (D) of ChE, human serum albumin, alpha- and gamma -globulins. The CCC separation was performed with optimized polymer phase systems in two different coiled columns. The best purification was achieved with 16.0% PEG 600-12.5% potassium phosphate at pH 9.2, by eluting the lower phase at 0.5 mL/min through the small capacity column in which ChE was purified directly from human serum within 2h. The ChE fractions confirmed by the enzymatic activity were free of serum proteins by SDS PAGE analysis.