Chromatography for Laboratory Instruction and Undergraduate Research

也许用于生物分子分析的最常见和最强大的分析技术是高效液相色谱（HPLC）。随着近年来生物技术与生物化学和化学的许多方面的整合，特别是HPLC，反相液相色谱法在蛋白质的纯化和表征中发挥着越来越重要的作用。通过这个项目，我们购买了两套高效液相色谱系统，主要用于高级本科生物化学和化学实验室。它们也可用于仪器分析课程和本科生独立研究项目。高效液相色谱系统正在生物化学实验室与毛细管电泳和十二烷基硫酸钠聚丙烯酰胺凝胶电泳一起使用，以分析过量产生重组蛋白的大肠杆菌细胞系的细胞提取物和纯化蛋白。一般来说，学生在本科实验室提供单一的方法来说明一个特定类型的分析。很少要求他们直接比较各种方法，以确定每种方法的相对优缺点。这些新装置被用于本科化学实验室，以评估不同流速和梯度剖面对一系列蛋白质洗脱模式的影响。学生将从理论和实践的角度考察分析化学家处理生物系统时所遇到的特殊考虑和困难。＊

Perhaps the most common and powerful analytical technique being used for the analysis of biomolecules is high-performance liquid chromatography (HPLC). With the recent integration of biotechnology into numerous aspects of biochemistry and chemistry, HPLC specifically, reversed-phase liquid chromatography is playing an increasingly more important role in the purification and characterization of proteins. Through this project, two HPLC systems are being purchased to be used primarily in the advanced undergraduate biochemistry and chemistry laboratories. They are also being made available both for use in the instrumental analysis class and for undergraduate independent study projects. The HPLC systems are being used in the biochemistry laboratory in conjunction with capillary electrophoresis and sodium dodecyl sulfate polyacrylamide gel electrophoresis to analyze cell extracts and purified protein from E. coli lines that overproduce a recombinant protein. Generally, students in undergraduate laboratories are provided with a single method to illustrate a particular type of analysis. Rarely are they asked to directly compare methods to determine the relative strengths and weaknesses of each method. The new units are being used in the undergraduate chemistry laboratory to evaluate the effect of varying the flow-rate and gradient profile on the elution patterns of a series of proteins. Students are examining from a theoretical and practical point of view, special considerations and difficulties encountered by the analytical chemist dealing with biological systems. *