摘要
目的采用荧光定量PCR法检测注射用重组人干扰素α2b半成品中宿主基因组的残留DNA。方法选择重组人干扰素α2b工程菌的宿主菌23S核糖体RNA基因为模板设计扩增引物,建立基于SYBR GreenⅠ荧光染料的荧光定量PCR检测法。结果宿主基因组DNA的量与荧光定量反应的Ct(循环阈值)值呈良好的线性关系(r2=0.9803)。结论所用方法操作简便、快速,可用于重组人干扰素α2b制备过程中的质量监控及半成品的检定。
OBJECTIVE To establish a method to determine residual host genomic DNA in recombinant human interferon α2b by real-time PCR.METHODS To design primers target to 23S ribosome RNA of host bacteria and amplify residual host genomic DNA of recombinant human interferon α2b by real-time PCR using SYBR GreenⅠ.RESULTS The concentration of residual host genomic DNA and the magnitude of cycle threshold(Ct) showed a good linear relationship.The correlation coefficient of standard curve was 0.9803.CONCLUSION The experim...
出处
《华西药学杂志》
CAS
CSCD
北大核心
2010年第2期166-168,共3页
West China Journal of Pharmaceutical Sciences
