PREPARATIVE CHROMATOGRAPHY OF PROTEINS

高效液相色谱(Hplc)已被证明是一种极好的 医学上重要的蛋白质和多肽的提纯和定量方法 例如变异和糖化血红蛋白、乳酸脱氢酶， 干扰素和胰岛素。不幸的是，现有的高效液相色谱柱 分析性质，不能制备性提纯克或 公斤合成或粗蛋白质，以满足其所需的质量 在人类身上的潜在用途。本申请描述了一种 专门设计的键合相液相色谱支架系列 用于蛋白质和多肽的大规模制备性纯化。 这样的载体将具有高容量和高分辨率，但也 僵硬而稳定。 第二阶段的具体目标将是综合、评估和选择 用于制备疏水和离子交换的制备载体 层析，以开发方法来扩大合成程序，以 50公斤水平，并制定操作参数，使最大限度地 在大型柱分析过程中加载和解析并将成本降至最低 这些支撑物中。 因为这种支撑物目前不可用，而且不耐用 使用可压缩凝胶作为替代品，其必要性是毋庸置疑的。 生物技术和制药公司正在提纯蛋白质和 多肽将是主要的商业用途。

High performance liquid chromatography (HPLC) has proven to be an excellent method to purify and quantitate medically important proteins and peptides such as variant and glycosylated hemoglobins, lactate dehydrogenase, interferon and insulin. Unfortunately, existing HPLC columns are analytical in nature and are incapable of preparatively purifying grams or kilograms of synthetic or crude proteins to a quality demanded by their potential use in humans. This application describes the synthesis of a series of bonded phase liquid chromatography supports specifically designed for the large-scale preparative purification of proteins and peptides. Such supports would have high capacity and high resolution, yet also be rigid and stable. The specific aims of Phase II would be to synthesize, evaluate and select preparative supports for preparative hydrophobic and ion-exchange chromatography, to develop methods to scale the synthetic procedures to 50kg levels, and to develop operational parameters which would maximize loading and resolution and minimize cost during analyses on large columns of these supports. Because such supports are unavailable at the present time and nondurable compressible gels are used as substitutes, the need is unquestionable. Biotechnological and pharmaceutical firms who are purifying proteins and peptides would be the major commercial users.