Earticle

초록

영어

Separation and/or removal of RNA and endotoxins, the major contaminants found in alkaline cell lysate in plasmid DNA (pDNA) purification process, has been cumbersome due to their similar properties (mostly negative charge) to pDNA. In our previous study, differential affinity of pDNA, RNA and endotoxins towards various metal ions was demonstrated that endotoxins and RNA preferentially and strongly bind to Cu2+-IDA and that pDNA binds only to Fe3+-IDA in the solution lacking RNA and endotoxins. By harnessing these preferential affinities in a single stage monolith-based immobilized metal affinity chromatography (IMAC) comprising Cu2+-IDA followed by Fe3+-IDA columns in series, pDNA was recovered in pure form at a high yield from the flowthrough from the Cu2+-IDA column where RNA and endotoxins were selectively captured. Furthermore, the proposed processing scheme, empowered by the advantages of monolith matrix (e.g. high flowrate and/or throughput, short processing time), exhibited significantly higher productivity over the conventional chromatographic processing schemes using particle based supports, facilitating the scale-up of the developed process to an industrial scale. Additional polishing step to remove genomic DNA and/or pDNA isoforms (e.g. linear and open circular forms) from supercoiled pDNA gave pharmaceutical-grade
pDNA capable of addressing regulatory specifications, thereby achieving significant intensification of pDNA purification process in an economically viable way.

키워드

저자

• Min-Jae SHIN [ School of Chemical Engineering, Sungkyunkwan University, Suwon, 440-746, Republic of Korea. ]
• Lihan TAN [ Department of Chemical and Biomolecular Engineering, National University of Singapore, 10 Kent Ridge Crescent, 117576 Singapore. ]
• Woo Seok CHOE [ School of Chemical Engineering, Sungkyunkwan University, Suwon, 440-746, Republic of Korea. ]

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576