Economic optimization of antibody capture through Protein A affinity nanofiber chromatography

Abstract

In this study we compare a nanofiber device and a resin column, both with the same Protein A ligand, for the capture of four different monoclonal antibodies and a trivalent crystallizable fragments (Fc) of antibody. We show that the dynamic binding capacity of the nanofiber device varies slightly with residence time, with the transfer kinetics slower for the IgG1, than for the IgG4 antibody. Surface plasmon resonance shows that the trivalent Fc binds to multiple Protein A ligand sites due to its three repeating Fc units, whereas multiple IgG antibodies bind to a single Protein A ligand. The nanofiber device gives comparable eluate purity to the resin column but at lower antibody concentrations due to its larger housing volume and lower binding capacity. It can be used for up to 200 cycles without performance loss and gives consistent eluate quality when scaled from 0.4 mL to 3.75 mL. We show that the use of a single nanofiber device to process one 2000 L clarified harvest through multiple cycles can reduce the Cost of Goods fivefold compared with a resin column. Costs are potentially lower for up to thirteen 2000 L harvests if device reuse can be validated. Nanofiber devices are thus an attractive option for manufacture of pre-clinical or clinical trial material where the number of batches is limited.