期刊
JOURNAL OF MOLECULAR RECOGNITION
卷 27, 期 4, 页码 184-189出版社
WILEY
DOI: 10.1002/jmr.2349
关键词
plasmid DNA; RNA; endotoxin; multimodal chromatography; hydrophobic interaction; ionic recognition
资金
- FCT, Portuguese Foundation for Science and Technology [SFRH/BD/47934/2008]
- Fundação para a Ciência e a Tecnologia [SFRH/BD/47934/2008] Funding Source: FCT
Multimodal chromatography is widely used for isolation of proteins because it often results in improved selectivity compared to conventional separation resins. The binding potential and chromatographic behavior of plasmid DNA have here been examined on a Capto Adhere resin. Capto Adhere is a recent multimodal chromatography material allowing molecular recognition between the ligand and target molecule, which is based on combined ionic and aromatic interactions. Capto Adhere proved to offer a very strong binding of nucleic acids. This property could be used to isolate plasmid DNA from a crude Escherichia coli extract. Using a stepwise NaCl gradient, pure plasmid DNA could be obtained without protein and endotoxin contaminations. The RNA fraction bound most strongly to the resin and could be eluted only at very high salt concentrations (2.0M NaCl). The chromatographic separation behavior was very robust between pH values 6 and 9, and the dynamic binding capacity was estimated to 60 mu g/ml resin. Copyright (c) 2014 John Wiley & Sons, Ltd.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据