Comparison of perfusion media and monoliths for protein and virus-like particle chromatography

• Perfusion media provide higher DBC for proteins compared to monolith.
• Perfusion model predicts effects of flow on DBC.
• Protein recovery upon elution is high for both perfusion media and monolith.
• VLP recovery low and flow direction dependent for monolith.
• VLP recovery behavior related to structural heterogeneity of monolith.

Structural and performance characteristics of perfusion chromatography media (POROS HS 20 and 50) and those of a polymethacrylate monolith (CIM SO3-1 tube monolith column) are compared for protein and virus-like particle chromatography using 1 mL columns. Axial flow columns are used for POROS while the monolith has a radial flow configuration, which provides comparable operating pressures. The POROS beads contain a bimodal distribution of pore sizes, some as large as 0.5 μm, which allow a small fraction of the mobile phase to flow within the particles, while the monolith contains 1–2 μm flow channels. For proteins (lysozyme and IgG), the dynamic binding capacity of the POROS columns is more than twice that of the monolith at longer residence times. While the DBC of the POROS HS 50 column decreases at shorter residence times, the DBC of the POROS HS 20 column for IgG remains nearly twice that of the monolith at residence times at least as low as 0.2 min as a result of intraparticle convection. Protein recoveries are comparable for all three columns. For VLPs, however, the eluted peaks are broader and recovery is lower for the monolith than for the POROS columns and is dependent on the direction of flow in the monolith, which is attributed to denser layer observed by SEM at the inlet surface of the monolith that appears to trap VLPs when loading in the normal flow direction.