Protein A chromatography remains the crucial step in mAb purification because of the high binding specificity and impurity clearance. In recent years, highly productive membrane adsorbers emerged as an alternative to traditional resins allowing for rapid purification of biomolecules. In this study, we tested three commercially available protein A membranes (Sartobind® Rapid A, HiTrap Fibro™ PrismA and GORE™ Protein Capture Device) regarding flow distribution, permeability and binding performance. As an application study using a cell-culture supernatant (CCS) containing monoclonal antibodies (mAbs), acidic and high pH wash steps were investigated regarding recovery and impurity removal. All membranes proved their applicability as highly productive capture media leading to high HCP and DNA removal with no observable influence on recovery. GORE™ Protein Capture Device exhibited a superior flow distribution but revealed diffusional limitations at high flow rates. Sartobind® Rapid A and HiTrap Fibro™ PrismA showed binding capacities of ∼ 40 g/L even at residence times (RTs) < 12 s but were limited by hydrodynamics suggesting room for improvement with optimized membrane housing.
Keywords: High pH wash; Membrane chromatography; Protein A affinity.
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