Earticle

초록

영어

Potential advantages of plasmid DNA (pDNA) vaccine over the conventional vaccines include higher stability and life-long immunity against multiple diseases in a single inoculation. In order to develop a scalable intensified process capable of selectively purifying pDNA from its major, difficult-to-remove contaminants such as RNA and endotoxin, we compared two chromatography platforms based on packed-bed column (PBC) and monolith column (MC). We first investigated the dynamic binding capacity (DBC) of RNA, a major contaminant following selective removal of endotoxin, to Cu2+-chelated IDA in PBC and MC using pure RNA solution at various concentrations (0.05 – 8 mg-RNA/ml) and flowrates (0.5 – 3 ml/min for PBC and 0.5 –7 ml/min for MC). RNA exhibited largely concentration-dependent increasing DBC profiles
prior to reaching a plateau (max DBC of 1.7 and 1.4 mg-RNA/ml for PBC and MC, respectively) above 0.5 mg/ml of RNA in the flowrate ranges investigated. It was confirmed that the DBC of RNA remained unchanged with the use of feedstock (partially purified from alkaline cell lysate) containing RNA and pDNA. Since PBC or MC has its own merit in view of binding capacity (i.e. higher maximum DBC for PBC) or throughput (i.e. higher flowrate for MC), the overall efficiency of each PBC- or MC-based process is determined by productivity defined as processed feedstock volume per unit resin volume per unit time. For several feedstocks with varying concentrations of pDNA and RNA, the productivity changed in an RNA concentration dependent manner, but generally MC showed higher productivities than PBC. This study proves that the use of monolith chromatographic support which has lower mass transfer limitation and higher throughput than those of conventional packed-bed supports has the benefits in processing macromolecules such as RNA and pDNA, thereby providing a more economically viable platform for pDNA purification.

키워드

저자

• Min Jae SHIN [ Department 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. ]
• Ik Keun YOO [ School of Chemical Engineering and Bioengineering, University of Ulsan, Ulsan 680-749, Republic of Korea. ]
• Woo Seok CHOE [ Department 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