Detergent-induced antibody elution at near-neutral pH in protein A membrane chromatography

Protein A affinity chromatography is the benchmark for monoclonal antibody (mAb) capture but typically requires low-pH elution (pH 3-4), which can compromise product stability. We evaluated a recombinant Protein A (rProtein A) ligand on a membrane platform, which supports higher baseline elution pH (4-5). Adding a virus inactivation detergent, an alkyl-chain amine oxide, further increased elution pH to a near-neutral range (5.5-6.5), mitigating risks of aggregation and activity loss. At pH 5.5, control elution conditions yielded 29.2 % product recovery, whereas addition of 1 % Deviron® C16 increased recovery to 99.5 %. Notably, recovery remained above 90 % at pH 6, whereas no elution occurred in the absence of Deviron® C16. Even when the detergent was introduced and removed prior to elution-applied only in an intermediate wash step-the recovery (86.9 %) exceeded the control at pH 5.5. The effect was detergent-specific, consistent across excipient concentrations, elution modes and immobilization format (membrane vs. column). Moreover, modulation varied across commercial resins bearing different engineered variants of the Protein A ligand, indicating dependence on ligand modifications. These results highlight an excipient-driven approach for gentler mAb capture and suggest the potential to integrate virus inactivation directly within the affinity purification step.