A continuous cell alkaline lysis, neutralization, and clarification combination process for production of plasmid pUDK-HGF |
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Authors: | Chunsheng Hu Qinglin Zhang Yuxin Lu Xiaochen Cheng Yanliang Wang Tong Zhang Zuze Wu |
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Institution: | Department of Experimental Hematology, Beijing Institute of Radiation Medicine, People's Republic of China. |
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Abstract: | 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 T-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. |
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