【摘要】 在昆虫细胞杆状病毒中表达的人生长激素 ,经 CM-5 2离子交换柱层析和 Phenyl-Sepharose CL-4 B疏水柱层析后 ,去除了 93%杂蛋白 .再用 CNBr-activated Sepharose4 B亲和柱层析的方法 ,获得高纯度的生长激素样品 .SDS-PAGE结果表明 ,纯化样品在 SDS-PAGE图谱呈现一条带 ,分子量为 2 2 k D.Western杂交结果表明 ,纯化样品与生长激素标准品有相似的免疫活性 .胰肽图谱分析结果显示纯化样品和标准品具有相同的一级结构 .DNS-CL测 N末端表明 ,纯化样品的 N末端为苯丙氨酸 ,所有这些性质均与天然人生长激素相同更多还原

【Abstract】 A protocol using Ion Exchange Chromatography (IEC), Hydrophobic Interaction Chromatography (HIC) and Affinity Chromatography for purifing recombinant human Growth Hormone (rhGH) expressed in insect cells was reported. IEC and HIC was used as the initial step for concentration and partial purification. The rhGH was extracted from insect cell culture fluid by batch absorption to the series-connected columns (CM-52 and Phenyl Sepharose CL-4B) in sequence. This procedure rendered a concentrated solution. The following step was Affinity Chromatography, which shows high efficiency of purification and recovery. The purified rhGH was determined to have identical sequence with native one by Tryptic peptide map; and also have identical immuitive activity by Western blotting; molecular weight of 22,000 daltons by SDS-PAGE; isoelectric point of pH 4.9 by isoelectric focus; Phe as N-terminal amino acid by DNS-CL terminal assaying method. All the properties conform with those of native hGH.更多还原