Biporous polymeric microspheres coupled with mercaptopyridine for rapid chromatographic purification of plasmid DNA

被引：0
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作者：
Li, Yuan [1 ]
Dong, Xiao-Yan [1 ]
Sun, Yan [1 ]
机构：
[1] Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China
来源：
Journal of Applied Polymer Science | 2007年 / 104卷 / 04期
关键词：
A biporous absorbent coupled with mercapto-pyridine was synthesized for the purification of plasmid DNA. Analyses by scanning electron microscopy and mercury intrusion porosimetry revealed that the matrix contained two families of pores; i.e; micropores and superpores. The super-pores provided not only convective flow channels for the mobile phase; but also a large surface for biomacromolecules binding. So the chromatographic process can be operated at high flow rate with high column efficiency and low backpressure as identified on a 2-mL column. When 10 mL crude feed-stock containing 3 mg of plasmid (5.4 kb pcDNA3) was loaded at a flow rate as high as 20 cm/min; the separation was finished in 10 min; and the plasmid was completely recovered with undetectable impurities of nucleic acids and proteins. The productivity was determined to be 9.0 g/L h; comparable to the pDNA productivity obtained using the commercial column. These results indicate that the biporous medium is promising for high-throughput purification of plasmid DNA. © 2007 Wiley Periodicals; Inc;
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页码：2205 / 2211
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