earticle

영어

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.