PREPARATIVE CHROMATOGRAPHY OF PROTEINS

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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.