16S rRNA gene probe quantitates residual host cell DNA in pharmaceutical-grade plasmid DNA

被引：8

作者：

Wang, KY ^{[1
]}

Guo, YJ ^{[1
]}

Sun, SH ^{[1
]}

Shi, K ^{[1
]}

Zhang, S ^{[1
]}

Wang, KH ^{[1
]}

Yi-Zhang ^{[1
]}

Chen, ZH ^{[1
]}

机构：

[1] Second Mil Med Univ, Dept Med Genet, Shanghai 200433, Peoples R China

来源：

VACCINE | 2006年 / 24卷 / 14期

关键词：

DNA vaccine; 16S rRNA; E. coli DNA;

D O I：

10.1016/j.vaccine.2005.11.066

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

The development and widespread use of DNA-based vaccination against infectious pathogens have been a great triumph of medical science. Quality control of DNA vaccines as biopharmaceutical productions is a problem to solve. Residual genomic DNA of engineering bacteria has been identified as a potential risk factor, so whose level must be controlled under the regulatory standards. We report a dot-blot hybridization method to detect residual host cell DNA in purified DNA vaccines. The assay utilizes PCR amplified and digoxigenin-labeled Escherichia coli 16S rRNA gene as probe. The sensitivity of the dot-blot hybridization assay with E. coli 16S rRNA gene probe was evaluated in comparison with single copy UidR gene probe. The optimized dot-blot hybridization assay had both low background and a suitable sensitivity, detecting 10 pg of residual E. coli DNA. The method is suitable in the routine use of measuring the levels of residual E. coli DNA in the pharmaceutical-grade DNA vaccine. (c) 2005 Elsevier Ltd. All rights reserved.

引用

页码：2656 / 2661

页数：6