【作者】 应跃斌； 潘映秋； 张伟； 陈枢青；

【Author】 YING Yue-bin, PAN Ying-qiu,ZHANG Wei, CHEN Shu-qing (Department of Biopharmaceutical, College of Phamaceutical Sciences, Zhejiang University. Hangzhou, Zhejiang 310006)

【机构】 浙江大学药学院生物制药研究室；

【摘要】 金黄色葡萄球茵肠毒素SEC2是超抗原家族的主要成员之一,已被证明为一临床药物的主要有效成分,对于肿瘤的免疫治疗有着良好的应用前景;以其为抗原而获得的单克隆抗体在食品、卫生检验领域也存在广阔的应用空间．本研究通过将产毒培养后的发酵液经超滤浓缩、硫酸铵沉淀、DEAE-Sepharose阴离子交换层析和SephdexG-75分子筛层析,成功地获得了高纯度的肠毒素SEC2．产物在SDS-PAGE上呈现单一条带,经光密度扫描分析,目的蛋白纯度大于90％, 回收率约为23％．体外刺激脾淋巴细胞增殖实验以及肿瘤杀伤实验表明该蛋白具备超抗原的活性．更多还原

【Abstract】 The Staphylococcal aureus enterotoxin SEC2 is an important one of superantigen family, and was proved to be the main effect element of a clinical drug. There is a wide application of SEC2 in the anticancer field, and the monoclone antibody for SEC2 is also applied in food test and clinical diagnoses. A method was developed to purify SEC2 from the broth of Staphylococcal aureus: after condensed by ultrafiltration, the protein was salt out by sulfate ammonium, the SEC2 fraction was isolated and purified by DEAE-Sepharose ion-exchange and G-75 molecular exclusion chromatography. The target protein purity was analysised by SDS-PAGE and densitometric scan to be more than 90%,and recover ratio is about 20%. The superantigen activity of the protein was proved by lymphocyte proliferation test and tumor suppression test.更多还原