色谱

色谱 ›› 2012, Vol. 30 ›› Issue (05): 522-526.DOI: 10.3724/SP.J.1123.2011.11045

• 研究论文 • 上一篇    下一篇

质粒DNA在灌注色谱基质上的吸附行为

Miladys LIMONTA1*, Lourdes ZUMALACRREGUI2, Dayana SOLER1   

  1. 1. Center for Genetic Engineering and Biotechnology, Habana 10600, Cuba; 2. Higher Polytechnic Institute José Antonio Echeverría Cuba, Habana 19390, Cuba
  • 收稿日期:2011-11-24 修回日期:2012-01-08 出版日期:2012-05-28 发布日期:2012-06-07
  • 通讯作者: Miladys LIMONTA, PhD, Associate professor. Tel: +53-7250-4432, E-mail: miladys.limonta@cigb.edu.cu.

Adsorption behavior of plasmid DNA onto perfusion chromatographic matrix

Miladys LIMONTA1*, Lourdes ZUMALACRREGUI2, Dayana SOLER1   

  1. 1. Center for Genetic Engineering and Biotechnology, Habana 10600, Cuba; 2. Higher Polytechnic Institute José Antonio Echeverría Cuba, Habana 19390, Cuba
  • Received:2011-11-24 Revised:2012-01-08 Online:2012-05-28 Published:2012-06-07
  • Contact: Miladys LIMONTA, PhD, Associate professor. Tel: +53-7250-4432, E-mail: miladys.limonta@cigb.edu.cu.

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摘要： Anion exchange chromatography is the most popular chromatographic method for plasmid separation. POROS R1 50 is a perfusion chromatographic support which is a reversed phase matrix and is an alternative to conventional ones due to its mass transfer properties. The adsorption and elution of the pIDKE2 plasmid onto reversed phase POROS R1 50 was studied. Langmuir isotherm model was adjusted in order to get the maximum adsorption capacity and the dissociation constant for POROS R1 50-plasmid DNA (pDNA) system. Breakthrough curves were obtained for volumetric flows between 0.69-3.33 mL/min, given dynamic capacity up to 2.3 times higher than those reported for ionic exchange matrix used during the purification process of plasmids with similar size to that of pIDKE2. The efficiency was less than 45% for the flow conditions and initial concentration studied, which means that the support will not be operated under saturation circumstances.

关键词: adsorption isotherm, perfusion chromatography, plasmid purification

Abstract: Anion exchange chromatography is the most popular chromatographic method for plasmid separation. POROS R1 50 is a perfusion chromatographic support which is a reversed phase matrix and is an alternative to conventional ones due to its mass transfer properties. The adsorption and elution of the pIDKE2 plasmid onto reversed phase POROS R1 50 was studied. Langmuir isotherm model was adjusted in order to get the maximum adsorption capacity and the dissociation constant for POROS R1 50-plasmid DNA (pDNA) system. Breakthrough curves were obtained for volumetric flows between 0.69-3.33 mL/min, given dynamic capacity up to 2.3 times higher than those reported for ionic exchange matrix used during the purification process of plasmids with similar size to that of pIDKE2. The efficiency was less than 45% for the flow conditions and initial concentration studied, which means that the support will not be operated under saturation circumstances.

Key words: adsorption isotherm, perfusion chromatography, plasmid purification