Grafting Chromatographic Monoliths With Charged Linear Polymers for Highly Productive and Selective Plasmid DNA Purification

Increased requirements for plasmid DNA (pDNA) in gene therapy and vaccination efforts brings the need for efficient large-scale production processes with high purity and productivity. Chromatographic purification of pDNA is often a bottleneck due to low dynamic binding capacities (DBC), pDNA recovery and/or low selectivity of columns. Ion exchangers (IEX) with charged groups on surface extenders (grafted and surface modified polymer chains formed by grafting reaction) have attracted attention due to their greatly enhanced productivity compared with traditional IEX.

The goal of our investigation was to increase the DBC for pDNA on a monolith stationary phase by grafting the surface with linear polymethacrylate brushes, while retaining high pDNA recovery and chromatographic selectivity between pDNA and RNA impurities. The effect of graft length and density on DBC and recovery was investigated. The optimal grafted column (with 1 mL bed volume) was evaluated for pDNA purification from neutralized E. coli lysate for two pDNA sizes, 4.7 and 11.6 kbp. Performance and possible scale-up of the grafted monolithic column for pDNA purification was presented with productivity calculation for anion-exchange (AEX) capture step.

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