SEPARATION OF RECOMBINANT PROTEINS BY VES ICLE CHROMATOGRAPHY

Vesicle chromatography, a recently developed method for separation of biomolecules, uses the vesicular packing (VP) material (clusters of mi crocapsules derived from plant cells), which was tested with respect t o its application for the recombinant protein separation. Since VP has a well-defined separation limit, biomolecules are distributed in two separate peaks: large molecules are excluded and small molecules perme ate through cell walls into the empty cell lumen. Recombinant proteins frequently form oligomers, which differ from monomers not only in siz e but also chemically and biologically. In the present study, separati ons of the recombinant proinsulin fusion protein oligomer and monomer, the recombinant human gamma-interferon monomer and dimer and recombin ant tumour necrosis factor-alpha were investigated. For peak identific ation, the fractions and starting samples of the recombinant proteins were analysed by HPLC. The separations occurred without any sorption e ffects and with high efficiency and resolution of the protein peaks at a short column (10 cm). The VP is characterised by a high load abilit y, which favours the scale-up purification of the recombinant proteins . The combination of VP and HPLC is a considerable advance in biotechn ology separation.