Experimental characterization of the transport phenomena, adsorption, and elution in a protein A affinity monolithic medium

• The advantage of convective media with HETP independent on flow rate is proved.
• Monolith bed utilization outperforms packed beds at low residence times.
• Adsorption of IgG is performed consistently in staircase and frontal analysis mode.
• Elution of IgG is limited by the kinetics of desorption at high pH (>4.0).
• At low pH (<4.0), convection is the limiting transport phenomenon during elution.

A commercially available convective interaction media (CIM) Protein A monolithic column was fully characterized in view of its application for the affinity capture of IgG in monoclonal antibody production processes. By means of moment analysis, the interstitial porosity and axial dispersion coefficient were determined. The frontal analysis method of characteristic points was employed, for the first time with monolithic media, to determine the dynamic binding capacity. The effects of the flow rate and pH on the total recovery of polyclonal IgG and elution profile were evaluated. A comparison with literature data for Protein A chromatography beads demonstrate the superior bed utilization of monolithic media, which gave better performance at lower residence times.