实时定量PCR法检测生物技术药物中宿主基因组DNA残留  

Detecting Residual Host Genomic DNA in Biotech Drugs by Real-Time Quantitative PCR

作　　者：曹晨华[1,2] 刘晓志[1,2] 段月娇 刘素霞 赵伟[1,2] 常亮 纪丽曼[1,2] 高健 

机构地区：[1]华北制药集团新药研究开发有限责任公司 [2]抗体药物研制国家重点实验室,石家庄050015

出　　处：《生物技术进展》2014年第2期142-145,共4页Current Biotechnology

基　　金：国家863计划项目(2012AA02A306);国家科技重大专项项目(2012ZX09101313)资助

摘　　要：利用基于SYBR GreenⅠ荧光染料的实时定量PCR方法检测酵母表达生物技术药物产品中宿主DNA残留量。该方法检测灵敏度可达到1.0 fg/μL,DNA浓度在1.0 fg/μL^1.0 ng/μL范围内线性良好,其标准曲线的相关系数为0.99以上。应用该方法对3批不同实验样本进行测定,宿主DNA残留量分别为8.635×105fg/μL、6.265×102fg/μL和1.436 fg/μL。实验表明该方法操作简便、灵敏度高,可用于生物技术药物产品中酵母DNA残留的定量测定。We detected residual host DNA in yeast expressing biotech drugs by SYBR Green I fluorescent based real-time quantitative polymerase chain reaction. The detection sensitivity of this method was up to 1.0 fg/μL. The linear correlation eoeflicient of standard eurve was above 0.99 when DNA concentration was bwtween 1.0 fg/μL to 1.0 ng/μL. Three batch of samples was determined, and the residual host DNA were 8.635× 105 fg/μL,6.265- 102 fg,/μL and 1.436 fg/μL respectively. The results suggested that this easy-handled, rapid and highly sensitive method could be used for the determination of residual yeast DNA in biotech drug products.

关 键 词：实时定量PCR 宿主基因组DNA残留 生物技术药物 质量控制 

分 类 号：R96[医药卫生—药理学]