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Comparison of Binding Capacity and Affinity of Monoclonal Antibody towards Different Affinity Resins using High-throughput Chromatography Method

Keywords: monoclonal antibody , high-throughput chromatography , Affinity resin , adsorption isotherm

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Abstract:

Protein-A Affinity chromatography is the widely used key method for purification of monoclonal antibodies. Selection of a most suitable affinity resin based on binding capacity and affinity is typically performed prior to optimization. Development of high-throughput chromatography method in 96-well filter plate significantly reduced consumption of antibody sample and shortens the experimental time as compared to a typical column chromatography approach. In this study, five different affinity resins were evaluated, rProtein-A FF, MabSelect Sure, ProSep-vA Ultra and two novel synthetically derived affinity ligands immobilized on agarose media, the GF1 and GF2 resins. Resins were dispensed on a 96-well filter plate and antibody sample with different protein concentration was loaded to evaluate resins affinity and static binding capacity. MabSelect Sure, an agarose based matrix with alkaline resistance Protein-A ligand and ProSep-vA Ultra that is a rigid pore glass resin exhibit the highest static binding capacity at ~60-63 mg IgG mL-1 of resin. The two novel resins, GF1 and GF2 show moderate binding capacity at ~28-34 mg IgG mL-1 of resin. By addition of salts during binding, the capacity of the novel resins was enhanced to ~33-42 mg IgG mL-1 resin. Affinity of all evaluated resins was quite comparable. Few other factors for resin selection such as dynamic binding capacity, ligand stability and resistance including resin cost will be briefly discussed.

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