A Mixed-Mode Approach — Alternative to a Three-Step mAb Purification Process

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Monoclonal antibody (mAb) purification methods often involve the use of resin-based chromatography in a series of steps. A typical mAb purification process involves a multi-step workflow consisting of two or more columns for capture, intermediate, and polish purification to isolate the target molecule. Resins selected for each of these steps solve specific challenges that exist at that phase of purification for the mAb. Purification strategies with combined mechanisms of action can allow for fewer chromatography steps to save processing time, thereby making enhancements to overall process economics and efficiency. We compare a three-step mAb purification using Nuvia Ion Exchange Resins post Protein A capture, to a two-step purification workflow with a Protein A capture step, followed by a mixed-mode chromatography step using Nuvia aPrime 4A, a hydrophobic anion exchange resin. We will present an additional two-step purification approach using Protein A Resin followed by a calcium affinity cation exchange media, CHT Ceramic Hydroxyapatite XT. The results of both studies will show the ability to achieve high purity product with one fewer unit operation.