【摘要】 高速逆流色谱是一种连续液-液色谱技术,具有无固相载体、样品无需严格预处理等优点。近10年来,在设备结构和溶剂体系等方面进行了大量的研究开发,已推广应用于生物技术、医药、天然产物、环境监测、食品等领域。为适应生物大分子和活性细胞的分离,采用条件温和的双水相体系,研究开发相应的高速逆流色谱设备已成为热点。针对双水相体系的特点,已经开发出了多种具有较高固定相保留率的新型高速逆流色谱设备,通过优化实验条件,成功地进行了多种蛋白质的分离纯化。本文对该领域的最新进展进行了综述与评价。更多还原

【Abstract】 High-speed countercurrent chromatography(HSCCC) is a kind of continuous liquid-liquid distribution/partition chromatography with the unique feature of eliminating the use of a solid support, which subsequently eliminates all complications arising therefrom, such as absorptive loss and denaturation of samples. In the past ten years, much research has been done concerning the development of apparatus and selection of solvent and promoted the applications of high speed countercurrent chromatography in various areas, especially the separation and preparation of natural products. However, the solvent systems used are mainly organic/aqueous systems, which tend to result in irreversible denaturalization of biological macromolecules. The aqueous two-phase system with mild operation conditions is preferred for separation of biomolecules. Yet the high viscosity and low interfacial tension of polymer phase tends to cause emulsification of the two phases and results in lower and unstable retention of stationary phase in conventional HSCCC apparatus. With the development of several types of novel high-speed countercurrent apparatus, including the cross-axis countercurrent chromatograph and the nonsynchronous countercurrent chromatograph, the retention of aqueous two-phase system in HSCCC has been greatly improved, which enables and highlights the countercurrent separation of biomolecules with aqueous two-phase systems. Various protein samples have been successfully separated in aqueous two-phase system with those newly-designed HSCCC apparatus. Recent advance in said area was summarized and evaluated in this paper.更多还原