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.
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
