Plasmid DNA (pDNA) vaccines have potential advantages over conventional vaccines, including higher safety and ability to induce long-lived immune response. The purification process for pDNA from impurities such as RNA and endotoxins in large-scale is required. The use of monolith chromatographic support which has higher porosity and throughput than those of conventional packed-bed support has the potential to be a new and efficient pDNA purification platform. We demonstrated that the use of Cu2+-IDA exhibited hierarchical preferential capture for endotoxins, RNA and pDNA in the decreasing order1. It was also shown that this hierarchical binding of pDNA and other impurities could be harnessed for developing a simple two-step pDNA purification process2. In the present study, we investigated the dynamic binding capacity of pDNA, RNA and endotoxins to Cu2+ chelated iminodiacetic acid (IDA) monolithic column. The dynamic binding capacity of RNA and endotoxins (determined from the frontal analysis of breakthrough curves) increased with the increase in RNA or endotoxins concentration, while that for pDNA is negligible. In addition, the breakthrough curves of RNA and endotoxins are independent of the flow-rate. These make the use of monolithic column ideal for large-scale processing at high concentration and flow-rate, rendering the purification process faster and more economically viable.
키워드
Plasmid DNA vaccine purificationMonolithIMACDynamic binding capacity
저자
Min Jae SHIN [ Dept. of Chemical Engineering, Sungkyunkwan University ]
Lihan TAN [ Dept. of Chemical and Biomolecular Engineering, National University of Singapore, 10 Kent Ridge Crescent, 117576 Singapore. ]
Woo Seok CHOE [ Dept. of Chemical Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea, Dept. of Chemical and Biomolecular Engineering, National University of Singapore, 10 Kent Ridge Crescent, 117576 Singapore. ]
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
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