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A continuous cell alkaline lysis, neutralization, and clarification combination process for production of plasmid pUDK‐HGF
Author(s) -
Chunsheng Hu,
Qinglin Zhang,
Yuxin Lu,
Xiaochen Cheng,
Yanliang Wang,
Tong Zhang,
Zuze Wu
Publication year - 2011
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.23
Subject(s) - lysis , alkaline lysis , plasmid , plasmid preparation , chromatography , escherichia coli , yield (engineering) , dna , downstream processing , chemistry , biology , microbiology and biotechnology , biochemistry , dna vaccination , materials science , gene , pbr322 , metallurgy
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.