PROCESS FOR PURIFYING HUMAN THROMBOPOIETIN WITH HIGH CONTENT OF SIALIC ACID

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.