An improved, inexpensive procedure for the large-scale purification of recombinant human erythropoietin.
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 160,104 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.
Hu
Yunlong
Y
Academy of Life Science, Nanjing Normal University, Nanjing 210094, People's Republic of China. ylhu@jlonline.com
Chen
Song
S
Xu
Mei
M
Zhang
Shuangquan
S
D000818:Animals; D005591:Chemical Fractionation / Q000379:methods*; D002845:Chromatography / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D006224:Cricetinae; D003412:Cricetulus; D004921:Erythropoietin / Q000302:isolation & purification* / Q000378:metabolism*; D006801:Humans; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins
D011994:Recombinant Proteins; D004921:Erythropoietin