title: A continuous cell alkaline lysis, neutralization, and clarification combination process for production of plasmid pUDK-HGF.
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.
authors.affiliation: Department of Experimental Hematology, Beijing Institute of Radiation Medicine, People's Republic of China.
authors.forename: Hu
authors.initials: H
authors.lastname: Chunsheng
authors.forename: Zhang
authors.initials: Z
authors.lastname: Qinglin
authors.forename: Lu
authors.initials: L
authors.lastname: Yuxin
authors.forename: Cheng
authors.initials: C
authors.lastname: Xiaochen
authors.forename: Wang
authors.initials: W
authors.lastname: Yanliang
authors.forename: Zhang
authors.initials: Z
authors.lastname: Tong
authors.forename: Wu
authors.initials: W
authors.lastname: Zuze
mesh_terms: D000468:Alkalies / Q000737:chemistry*; D001709:Biotechnology; D004926:Escherichia coli / Q000166:cytology* / Q000235:genetics*; D005285:Fermentation; D010957:Plasmids / Q000302:isolation & purification*
chemical_list: D000468:Alkalies