Preparative High Performance Capillary Electrophoresis

高性能毛细管电泳（HPCE）在蛋白质和核酸的表征方面比传统的凝胶电泳具有重要的优势。它比凝胶快得多，可以完全自动化，并具有样品注射，检测和收集的仪器化。此外，高效液相色谱分离溶质的分辨率比高效液相色谱（HPLC）高一到两个数量级，而样品大小比高效液相色谱（HPLC）小四个数量级。然而，单毛细管高效液相色谱只能处理小于1 ug的溶质负载。它不能用于预备规模，为进一步分析、临床研究或治疗应用提供材料。大多数熟悉这项技术的科学家认为，如果HPCE能够达到制备水平，它将成为不可或缺的实验室工具。不幸的是，放大到制备HPCE并不简单：制备HPCE柱必须有效地消散焦耳热，同时保持在ug中常规观察到的非常高的板效率。HPCE。为了达到1 mg高效毛细管电泳的目标，提出了一项将单毛细管和多毛细管高效毛细管电泳的理论和实验分析相结合的研究。本研究的目的是：(1)阐明单个毛细管中的传热、分离和分散机制，并将这些知识应用于(2)制备级高效液相色谱柱的设计。

High performance capillary electrophoresis (HPCE) offers important advantages over conventional gel electrophoresis in the characterization of proteins and nucleic acids. It is much faster than the gels and can be fully automated and instrumented with sample injection, detection and collection. In addition, HPCE can separate solutes with a resolution which is one to two orders of magnitude greater than high performance liquid chromatography (HPLC) using sample sizes four orders of magnitude smaller than HPLC. However, single-capillary HPCE can only handle solutes loadings of less than 1 ug. It cannot be used at preparatory scales to provide material for further analysis, clinical research or therapeutic application. Most scientists familiar with this technology believe that, if HPCE could be scaled to preparative levels, it would become an indispensable laboratory tool. Unfortunately, scale-up to preparative HPCE is not simple: a preparative HPCE column must dissipate Joule heat effectively while maintaining the extraordinarily high plate efficiency routinely observed in ug. HPCE. To reach the goal of 1 mg HPCE, a study is proposed which combines both theoretical and experimental analyses of single and multiple capillary HPCE. The objectives of this research are to (1) elucidate the mechanisms of heat transfer, separation and dispersion in single capillaries and to apply this knowledge in the (2) design of a preparative-scale HPCE column.