Departamento de Ingeniería Química y Metalurgia, Universidad de Sonora, Hermosillo, Sonora, México.
Biotechnol Appl Biochem. 2011 Jan-Feb;58(1):68-74. doi: 10.1002/bab.12.
A novel downstream bioprocess was developed to obtain purified plasmid DNA (pDNA) from Escherichia coli ferments. The intermediate recovery and purification of the pDNA in cell lysate was conducted using hollow-fiber tangential filtration and frontal anion-exchange membrane and elution hydrophobic chromatographies. The purity of the solutions of pDNA obtained during each process stage was investigated. The results show that the pDNA solution purity increased 30-fold and more than 99% of RNA in the lysate was removed during the process operations. The combination of membrane operations and hydrophobic interaction chromatography resulted in an efficient way to recover pDNA from cell lysates. A better understanding of membrane-based technology for the purification of pDNA from clarified E. coli lysate was developed in this research.
开发了一种新颖的下游生物处理方法，从大肠杆菌发酵液中获得纯化的质粒 DNA（pDNA）。通过中空纤维切向过滤、前沿阴离子交换膜和洗脱疏水性色谱法对细胞裂解物中的 pDNA 进行中间回收和纯化。研究了在每个工艺阶段获得的 pDNA 溶液的纯度。结果表明，在该过程操作过程中，pDNA 溶液的纯度提高了 30 倍，并且裂解物中超过 99%的 RNA 被去除。膜操作和疏水相互作用色谱法的结合为从细胞裂解物中回收 pDNA 提供了一种有效的方法。本研究开发了一种更好地理解基于膜的技术，用于从澄清的大肠杆菌裂解物中纯化 pDNA。
