| 一种简便易行核酸疫苗质粒纯化工艺的开发 |
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| 引用本文： | 张平静,李忠明,刘庆良.一种简便易行核酸疫苗质粒纯化工艺的开发[J].中国生物工程杂志,2011,31(4):106-112. |
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| 作者姓名： | 张平静 李忠明 刘庆良 |
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| 作者单位： | 上海海规生物科技有限公司 上海 201203 |
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| 基金项目： | 国家科技部"十一五"国家重大专项课题任务基金 |
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| 摘 要： | 介绍了一种成本低、步骤少、简单易行的质粒纯化制检工艺。该工艺选择优势产生超螺旋质粒的大肠杆菌菌株以无蛋白质培养基进行发酵罐培养,采用碱裂解法,对质粒制备过程中所用的层析吸附材料、核酸结合溶液、去除内毒素等杂质的方法和浓缩等步骤进行了实用性改进,并建立了相应的检定方法,所得质粒的纯度达到临床级要求。
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| 关 键 词： | 核酸疫苗 质粒 纯化 内毒素 硅胶 |
| 收稿时间： | 2010-09-08 |
| 修稿时间： | 2011-01-28 |
The Development of a Modified and Simple Process for the Purification of Plasmid DNA |
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| ZHANG Ping-jing,LI Zhong-ming,LIU Qing-liang.The Development of a Modified and Simple Process for the Purification of Plasmid DNA[J].China Biotechnology,2011,31(4):106-112. |
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| Authors: | ZHANG Ping-jing LI Zhong-ming LIU Qing-liang |
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| Institution: | (Shanghai H & Q Biotech Lid Cop,Shanghai 201203,China) |
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| Abstract: | A modified and simple manufacturing scale process for the purification of plasmid DNA was developed with low cost. The Escherichia coli JM108 host strain was selected for the fermentation with protein free media because of its advantage to produce predominantly super coiled plasmid DNA. The alkaline lyses method was used. The plasmid purification process including chromatography-absorbent silica, suitable DNA binding solution, endotoxin-removal reagents was substantially modified. The tests for the quality control were established for the detection of the residue impurities. The purity of the plasmid DNA obtained by this process can meet the clinical quality requirements. |
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| Keywords: | DNA vaccine Plasmid Purification Endotoxin Silica |
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