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.
Academy of Life Science, Nanjing Normal University, Nanjing 210094, People's Republic of China. ylhu@jlonline.com
Yunlong
Y
Hu
Song
S
Chen
Mei
M
Xu
Shuangquan
S
Zhang
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