Rapid purification of mAb using protein a membranes yielding high HCP clearance

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2024-01, Vol.1232, p.123989-123989, Article 123989
• Main Authors: Gehrmann, Nils, Daxbacher, Andreas, Hahn, Rainer
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - chemistry; CHO Cells; Chromatography; Chromatography, Affinity - methods; Cricetinae; Cricetulus; High pH wash; Kinetics; Membrane chromatography; Protein A affinity; Staphylococcal Protein A
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2023.123989

•GORE Protein Capture Device™ with enhanced flow distribution inside membrane device but limited by diffusion.•Sartobind Rapid A Nano™ highly permeable, only slightly limited by diffusion.•HiTrap Fibro PrismA™ with no loss of binding capacity at high flow rates.•Protein A membranes show excellent applicability in mAb purification.•Enhanced HCP removal using high pH wash, no trends regarding wash volume or flow rate. 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)