肿瘤基因疫苗纯化工艺的建立  

Development of Purification Procedure for Tumor DNA Vaccine

作　　者：杨晓玲[1] 吕婧[1] 朱智强[1] 谢秋[1] 王建华[1] 张悦红[1] 牛勃[1] 

机构地区：[1]山西医科大学生物化学与分子生物学教研室,太原030001

出　　处：《中国生物制品学杂志》2010年第6期623-626,共4页Chinese Journal of Biologicals

摘　　要：目的建立肿瘤基因疫苗的纯化工艺,为肿瘤基因疫苗的工业化生产奠定基础。方法采用碱裂解法粗提质粒DNA,利用凝胶过滤层析、亲和层析及离子交换层析分离纯化质粒DNA,并对所纯化的质粒DNA进行全面检定。结果经3种层析柱纯化所获得的超螺旋质粒DNA,其A260与A280的比值在1.85～1.92之间;超螺旋质粒DNA比例不低于94%;内毒素含量远低于10EU/mg;5批纯化的质粒DNA经全面检定,均符合国家质量标准。结论该纯化工艺制备的肿瘤基因疫苗纯度高,且有效去除了内毒素。Objective To develop a purification procedure and lay a foundation of industrial production of tumor DNA vaccine.Methods Plasmid DNA was crudely extracted by alkaline lysis,then purified by gel filtration,affinity and ion-exchange chromatography,and subject to overall control tests.Results The ratio of A260 to A280 of plasmid DNA after purification by three steps of chromatography was 1.85 ～ 1.92,in which the proportion of super-coiled plasmid DNA was not less than 94%,and the endotoxin content was far less than 10 EU /mg.Overall control tests proved that all the five batches of purified plasmid DNAs met the national quality standards.Conclusion The tumor DNA vaccine purified by the developed procedure showed high purity,in which the endotoxin was removed effectively.

关 键 词：肿瘤 基因 疫苗 质粒 DNA 纯化 

分 类 号：R730.5[医药卫生—肿瘤]