【摘要】 对啤酒酵母生产的重组水蛭素变异体1(rHV1)进行多步骤的纯化。首先将分泌到培养上清液中的水蛭素进行硫酸铵沉淀和SephadexG-50凝胶过滤,再用Q-SepharoseHP阴离子交换层析分离,最后用HPLCSP-5PW阳离子交换柱脱色及HPLCC8柱反相层析。真空干燥后得到的水蛭素蛋白经SPS-PAGE、N端氨基酸序列分析、抗凝血酶活力分析鉴定,证明已获得高纯度的重组水蛭素HV1制剂,为利用基因工程方法生产重组水蛭素的规模化生产及临床应用提供了依据更多还原

【Abstract】 Recombinant hirudin produced by Saccharomyces cerevisiae was highly purified Hirudin expressed by yeast and secreted to culture supernatant was first precipitated by(NH 4) 2SO 4 and filtrated by Sephadex G 50 Then it was isolated by Q Sepharose HP anion exchange chromatography Last it was decolourized by HPLC cation exchange chromatography of SP 5PW column After reverse phase HPLC of C8 column and vacuum drying,the purely white hirudin protein was obtained.It was proved to be correct recombinant hirudin HV1 by SDS PAGE,amino acid sequence analysis of N terminal and activity identification of antithrombin The results supplied evidence for scale production and clinical application of recombinant hirudin produced by gene engineering method更多还原