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A Simple Method for Analyzing Aggregates of EPO (Erythropoietin) Using a BioSep™-SEC-s2000 GFC Column

Phenomenex Inc
30 Apr 2012

In this application note by Phenomenex, a rugged yet simple method is presented for separating EPO and its dimer. The difference in the amounts of aggregate present in both fresh and long-term stored EPO samples is determined using the BioSep-SEC-s2000 column. Recombinant EPO is approximately 30 kDa molecular weight in its glycosylated form (approximately 18 kDa for proteins), and any dimer of EPO would be expected to be around 60 kDa in size. A BioSep-SEC-s2000 series column was used for EPO separation by gel filtration chromatography, as it provides a large separation window for low molecular weight proteins.

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BioSep™ Aqueous GFC/SEC Columns

Phenomenex Inc

Economical, high efficiency columns to purify and/or characterize proteins, peptides, and other biomolecules including antibodies, aggregates, and PEGylated proteinsHigh performance: Analytical and preparative BioSep columns offer high resolution, maximum efficiency, and exceptional peak asymmetryImproved recovery: Extremely inert media offers advantages for accurte quantitationFlexible method development: 3 phases options (2000, 3000, 4000) to separate samples of varying molecular weight rangesPhenomenex offers support services worldwide through PhenoLogix, an in house analytical support laboratory. The team provides full support for labs needing method development and optimization support for new size excluion methods, as well as transferring current methods from other GFC media to BioSep-SEC-S phases.

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A Simple Method for Analyzing Aggregates of EPO (Erythropoietin) Using a BioSep™-SEC-s2000 GFC Column