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

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.