重组人血小板生成素的分离和纯化

Isolation and purification of recombinant human thrombpoietin

目的探讨重组人血小板生成素 (rhTPO)的分离、纯化工艺。方法高效表达rhTPO的CHO细胞的无血清培养液 ,经离心去除细胞碎片后 ,以二步液相色谱法进行纯化 ,对所得产物以HPLC法及SDS PAGE法进行纯度检测 ,用MO7e细胞 ,MTS细胞计数法测定其生物活性。结果终产品为单一条带、单一峰 ,生物活性为 0 .9× 10 5U/mg。结论经分离与纯化后得到的TPO产品 ,纯度高、生物活性好 。

PurposeThe aim is to explore isolation and purification of the Recombinant Human Thrombpoietin(rh TPO). MethodsUsing serum culture to obtain the culture fluid of high density and high rhTPO expressing. After centrifu ging, the residues of CHO cells were removed. We use two step chromatography procedure and get the product of rhTPO testing for its purity and bioactivity. Results1.Purity analysis: HPLC method and SDS PAGE method showed a single peak and a single band corresponded. Its purity surpassed by 96%. 2.Bioactivity assay: the bioactivity of this product was 0.9×10 5 IU/mg. ConclusionThe product of rhTPO had good repetition,and it′s suitable for large scale production.