Flux-dependent transmission of supercoiled plasmid DNA through ultrafiltration membranes
Flux-dependent transmission of supercoiled plasmid DNA through ultrafiltration membranes
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DOI：
10.1016/j.memsci.2007.02.033
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发表时间：
2007-05-15
影响因子：
9.5
通讯作者：
Zydney, Andrew L.
中科院分区：
文献类型：
--
作者：
Latulippe, David R.;Ager, Kimberly;Zydney, Andrew L.
尽管先前的研究已经证明了利用膜基工艺纯化质粒DNA的潜力，但在控制DNA传输的物理机制以及膜孔径和操作条件对DNA筛选系数的影响方面存在相当大的差异。这项工作的目的是获得3.0 kbp的超螺旋质粒DNA在一定滤液通量范围内通过不同标称分子量切断的Ultracel超滤膜的定量数据。质粒透过的程度与滤液通量有很强的关系，随着滤液通量的增加，筛分系数从基本为零增加到接近1。根据现有的过滤模型对数据进行分析，以检验DNA伸长、剪切变形和浓度极化对观察到的质粒传输的贡献。结果清楚地表明流动诱导的延伸或变形对DNA传输的重要性，数据与使用质粒超螺旋半径作为特征维的延长流动模型的改进版本一致。这些结果对控制DNA通过半透性超滤膜传输的因素提供了重要的见解。(C) 2007 Elsevier B.V.版权所有
Although previous studies have demonstrated the potential of using membrane-based processes for purification of plasmid DNA, there is considerable discrepancy regarding both the physical mechanisms governing DNA transmission and the effects of membrane pore size and operating conditions on the DNA sieving coefficient. The objective of this work was to obtain quantitative data on the transmission of a 3.0 kbp supercoiled plasmid DNA through Ultracel ultratiltration membranes with different nominal molecular weight cut-offs over a range of filtrate flux. The extent of plasmid transmission was a very strong function of the filtrate flux, with the sieving coefficient increasing from essentially zero to nearly one as the flux increased. Data were analyzed in terms of available filtration models to examine the contributions of DNA elongation, shear deformation, and concentration polarization on the observed plasmid transmission. The results clearly indicate the importance of flow-induced elongation or deformation on DNA transmission, with the data consistent with a modified version of the elongational flow model using the plasmid superhelix radius as the characteristic dimension. The results provide important insights into the factors controlling DNA transmission through semipermeable ultrafiltration membranes. (C) 2007 Elsevier B.V. All rights reserved.