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An improved, inexpensive procedure for the large‐scale purification of recombinant human erythropoietin
Author(s) -
Hu Yunlong,
Chen Song,
Xu Mei,
Zhang Shuangquan
Publication year - 2004
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20030189
Subject(s) - erythropoietin , recombinant dna , size exclusion chromatography , chromatography , yield (engineering) , chemistry , ion chromatography , specific activity , biochemistry , amino acid , high performance liquid chromatography , biology , enzyme , materials science , metallurgy , gene , endocrinology
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.