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质粒DNA在灌注色谱基质上的吸附行为。

Adsorption behavior of plasmid DNA onto perfusion chromatographic matrix.

作者信息

Limonta Miladys, Zumalacárregui Lourdes, Soler Dayana

机构信息

Center for Genetic Engineering and Biotechnology, Habana 10600, Cuba.

出版信息

Se Pu. 2012 May;30(5):522-6. doi: 10.3724/sp.j.1123.2011.11045.

Abstract

Anion exchange chromatography is the most popular chromatographic method for plasmid separation. POROS RI 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.

摘要

阴离子交换色谱法是质粒分离中最常用的色谱方法。POROS RI 50是一种灌注色谱载体,它是一种反相基质,由于其传质特性,是传统载体的一种替代物。研究了pIDKE2质粒在反相POROS R1 50上的吸附和洗脱。对朗缪尔等温线模型进行了调整,以获得POROS R1 50 - 质粒DNA(pDNA)系统的最大吸附容量和解离常数。在体积流速为0.69 - 3.33 mL/min的条件下获得了穿透曲线,其动态容量比在纯化与pIDKE2大小相似的质粒过程中使用的离子交换基质所报道的动态容量高出2.3倍。在所研究的流动条件和初始浓度下,效率低于45%,这意味着该载体不会在饱和条件下运行。

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