肝细胞生长因子
质粒
碱裂解
重组DNA
DNA
生物
基因
质体制备
溶解
分子生物学
遗传增强
离子色谱法
化学
生物化学
dna疫苗
受体
PBR322电话
作者
Qinglin Zhang,Jianjing Bi,Fengjun Xiao,Haifeng Duan,Bing Wu,Zhiyong Wu
摘要
pUDK-HGF, recombinant plasmid DNA encoding human HGF (hepatocyte growth factor), is a potential agent for gene therapy of ischaemic disease. Production of pUDK-HGF is essential for its clinical application. In the present paper, a large-scale manufacturing process was developed, including fermentation, cell harvest, alkaline lysis, capturing plasmid DNA with Q-Sepharose XL chromatography, size-exclusion chromatography on a Sephacryl S1000 column and refining with Source 15Q anion-exchange chromatography. The quality criteria of pUDK-HGF such as purity, concentration, homogeneity, residual RNA, chromosomal DNA, contaminated protein, endotoxin and HGF expression efficacy all were analysed and met the requirements for pharmaceutical-grade plasmid DNA.
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