Plasmid DNA partitioning and separation using poly(ethylene glycol)/poly(acrylate)/salt aqueous two-phase systems

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• 作者：Hans-Olof Johansson^a ; ^{Hans-Olof.Johansson@biochemistry.lu.se} ; Tiago Matos^b ; ^{tiago.matos@tbiokem.lth.se} ; Juliana S. Luz^c ; ^{jsluz.sp@gmail.com} ; Eloi Feitosa^d ; ^{eloifeitosa@gmail.com} ; Carla C. Oliveira^c ; ^{ccoliv@iq.usp.br} ; Adalberto Pessoa Jr.^e ; ^{pessoajr@usp.br} ; Leif Bü ; low^b ; ^{Leif.Bulow@tbiokem.lth.se} ; Folke Tjerneld^a ; ^{Folke.Tjerneld@biochemistry.lu.se}
• 关键词：Plasmid DNA ; Partitioning ; Aqueous two-phase systems ; Poly(ethylene glycol) ; Polyacrylate
• 刊名：Journal of Chromatography A
• 出版年：2012
• 期刊代码：47_00219673
• 类别：ch
• 出版时间：13 April, 2012
• 卷：1233
• 期：Complete
• 页码：30-35
• 文件大小：457 K

摘要

Phase diagrams of poly(ethylene glycol)/polyacrylate/Nab>2b>SOb>4b> systems have been investigated with respect to polymer size and pH. Plasmid DNA from Escherichia coli can depending on pH and polymer molecular weight be directed to a poly(ethylene glycol) or to a polyacrylate-rich phase in an aqueous two-phase system formed by these polymers. Bovine serum albumin (BSA) and E. coli homogenate proteins can be directed opposite to the plasmid partitioning in these systems. Two bioseparation processes have been developed where in the final step the pDNA is partitioned to a salt-rich phase giving a total process yield of 60-70%. In one of them the pDNA is partitioned between the polyacrylate and PEG-phases in order to remove proteins. In a more simplified process the plasmid is partitioned to a PEG-phase and back-extracted into a Nab>2b>SOb>4b>-rich phase. The novel polyacrylate/PEG system allows a strong change of the partitioning between the phases with relatively small changes in composition or pH.