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publicationDate 1995-03-22-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber EP-0228452-B1
titleOfInvention Protein purification
abstract Chromatographic procedures are individually and jointly applied to the rapid and efficient isolation of biologically active proteins and especially glycoproteins such as recombinant erythropoietin present in the medium of growth of genetically transformed mammalian host cells. Illustratively, recombinant EPO is isolated from culture fluids by reverse phase chromatography employing a C4 or C6 column and elution with ethanol. Recombinant erythropoientin may also be purified by anion exchange chromatography employing, e.g., a DEAE resin, with preliminary selective elution of contaminant materials having a lower pKa than erythropoietin from the resin under conditions mitigating against acid activated protease degradation. Practiced serially, the two chromatographic procedures allow for high yields of biologically active recombinant erythropoietin from mammalian cell culture media.
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