An improved, inexpensive procedure for the large‐scale purification of recombinant human erythropoietin

Abstract

A rapid and simple chromatographic procedure has been developed for the large‐scale purification of therapeutic‐grade rHuEPO (recombinant human erythropoietin) from medium‐conditioned cell cultures, which includes ion‐exchange, hydrophobic‐interaction and gel‐filtration chromatography. A combination of these well‐connected steps results in highly purified rHuEPO (>99%), as revealed by SDS/PAGE and HPLC analyses, with a total yield of 38%. The specific activity of purified rHuEPO was 160104 i.u./mg. Immunoblotting studies revealed that the protein possesses native EPO immunity. N‐terminal sequencing of rHuEPO shows that the first 15 amino acids coincide with those of native EPO reported previously.