Schmuck M N, Gooding D L, Gooding K M
J Chromatogr. 1986 May 30;359:323-30. doi: 10.1016/0021-9673(86)80086-3.
Although analytical high-performance liquid chromatographic columns have been successfully used for purification of milligram amounts of proteins, they do not appear to be ideal for preparing gram or kilogram quantities because of cost and load capacities. In this paper the development of preparative weak anion-exchange materials is described. These materials possess similar chromatographic characteristics to analytical 5-10 micron materials, yet also have high load capacities. A number of inorganic packings were examined to determine which had the best combination of high load capacity, good resolution, stability, and low cost. When appropriate flow-rates and gradient shapes were used, 30-50 micron materials produced resolution of components of a commercial ovalbumin sample that was comparable to that achieved on a 6-micron material. An amount of 3 g of a protein could be loaded onto a 250 X 21 mm-I.D. column with adequate resolution to separate it from some of its impurities.
尽管分析型高效液相色谱柱已成功用于毫克级蛋白质的纯化，但由于成本和负载能力的原因，它们似乎并不适合制备克级或千克级的蛋白质。本文描述了制备型弱阴离子交换材料的开发。这些材料具有与5 - 10微米分析型材料相似的色谱特性，但也具有高负载能力。研究了多种无机填料，以确定哪种填料具有高负载能力、良好的分离度、稳定性和低成本的最佳组合。当使用适当的流速和梯度形状时，30 - 50微米的材料对商业卵清蛋白样品各组分的分离度与6微米材料相当。3克蛋白质可以加载到内径为250×21毫米的柱上，并具有足够的分离度以将其与一些杂质分离。
