公开(公告)号：US20070264710A1

公开(公告)日：2007-11-15

申请号：US10573359

申请日：2003-10-09

申请人： Hyea-Kyung Ahn ,  Joo-Young Chung ,  Young-Ju Cho ,  Tae-Soo Kim ,  Yeo-Wook Koh ,  Seung-Wook Lim ,  Yeon-Joo Hong

发明人： Hyea-Kyung Ahn ,  Joo-Young Chung ,  Young-Ju Cho ,  Tae-Soo Kim ,  Yeo-Wook Koh ,  Seung-Wook Lim ,  Yeon-Joo Hong

IPC分类号： C07K1/00 ,  C12N5/06

CPC分类号： C07K14/524

摘要： Disclosed is a process for producing a human thrombopoietin (hTPO)-containing culture fluid, comprising the step of culturing a eukaryotic cell expressing hTPO in a serum-free medium that contains a negligible amount of serum. In addition, the present invention discloses a process for purifying hTPO from a hTPO-containing biological fluid, comprising the steps of (a) subjecting the biological fluid to affinity chromatography; (b) subjecting an eluate obtained at step (a) to hydrophobic interaction chromatography; (c) subjecting an eluate obtained at step (b) to reverse phased chromatography; and (d) subjecting an eluate obtained at step (c) to anion exchange chromatography. In addition, the present invention discloses hTPO with a high sialic acid content obtained by the process which comprises the step of loading, the eluate obtained at step (c) onto an ionic exchange chromatography column and collecting hTPO with a high content of sialic acid eluted selectively from the column by a 0.15-0.3M sodium chloride gradient.

摘要翻译： 公开了一种生产含有血小板生成素（hTPO）的培养液的方法，包括在含有可忽略量的血清的无血清培养基中培养表达hTPO的真核细胞的步骤。 另外，本发明公开了一种从含hTPO的生物流体中纯化hTPO的方法，包括以下步骤：（a）使生物液体进行亲和层析; （b）使步骤（a）获得的洗脱液进行疏水相互作用层析; （c）使步骤（b）获得的洗脱液反相定相; 和（d）使步骤（c）获得的洗脱液进行阴离子交换层析。 此外，本发明公开了通过该方法获得的具有高唾液酸含量的hTPO，其包括将步骤（c）获得的洗脱液装载到离子交换色谱柱上并收集高含量的唾液酸洗脱的hTPO 选择性地从柱中加入0.15-0.3M氯化钠梯度。