A continuous cell alkaline lysis, neutralization, and clarification combination process for production of plasmid pUDK-HGF

作者：Hu Chunsheng; Zhang Qinglin^*; Lu Yuxin; Cheng Xiaochen; Wang Yanliang; Zhang Tong; Wu Zuze

来源：Biotechnology and Applied Biochemistry, 2011, 58(3): 162-165.

DOI：10.1002/bab.23

• plasmid DNA
• alkaline lysis
• hollow fiber ultrafiltration column

Plasmid DNA for biopharmaceutical applications is produced easily in Escherichia coli bacteria. The cell lysis is the most crucial step for purification of plasmid DNA. In this paper, we describe a continuous cell alkaline lysis, neutralization, and clarification combination process for production of plasmid pUDK-HGF using hollow fiber ultrafiltration column as a lysis chamber and compare the plasmid DNA yield and homogeneity with the Y-connector and manual processes, respectively. The results show that the plasmid pUDK-HGF yield of the combination process is 13% higher than manual lysis, twice higher than using T-connector. When the proportion of lysed cells and neutralization solution is 3:1, the plasmid pUDK-HGF yield can improve by 70%. This process could be easily scaled up to meet the industrial scale for cell lysis.

• 出版日期2011-6
• 单位北京中医药大学; 内蒙古工业大学