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)。近年来，随着生物技术在生物化学和化学领域的广泛应用，反相液相色谱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，购买了两个高效液相系统，主要用于高等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的自主学习项目。生化实验室正在将高效液相系统与毛细管电泳法和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法结合使用，以分析过量生产重组蛋白的大肠杆菌菌株的细胞提取物和纯化蛋白。一般来说，本科实验室的学生只有一种方法来说明特定类型的分析。很少有人要求他们直接比较各种方法，以确定每种方法的相对优势和劣势。这些新的装置正被用于本科化学实验室，以评估改变流速和梯度曲线对一系列蛋白质洗脱模式的影响。学生从理论和实践的角度研究分析化学家在处理生物系统时遇到的特殊问题和困难。*

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. *