An improved, inexpensive procedure for the large-scale purification of recombinant human erythropoietin
An improved, inexpensive procedure for the large-scale purification of recombinant human erythropoietin
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DOI：
10.1042/ba20030189
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发表时间：
2004-08-01
影响因子：
2.8
通讯作者：
Zhang, SQ
中科院分区：
文献类型：
--
作者：
Hu, YL;Chen, S;Zhang, SQ
通过离子交换、疏水相互作用和凝胶过滤层析，建立了一种快速、简便的从培养细胞中大规模纯化治疗级重组人促红细胞生成素的层析方法。这些连接良好的步骤相结合，得到了高纯度的rHuEPO(>；99%)，如SDS/PAGE和H PLC分析所示，总收率为38%。纯化后的rHuEPO的比活力为160~104IU./mg。免疫印迹研究表明，该蛋白具有天然的EPO免疫力。对rHuEPO的N末端测序表明，第一个氨基酸与已报道的天然EPO的氨基酸序列一致。
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 H PLC 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 IS amino acids coincide with those of native EPO reported previously.