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

Monoclonal antibody (mAb) purification methods often involve the use of resin-based chromatography. A typical mAb purification process involves a multistep workflow using 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 mAb purification. Purification strategies with combined mechanisms of action can allow for fewer chromatography steps, saving processing time and thereby enhancing overall process economics and efficiency.

In this video, we compare a three-step mAb purification workflow, which uses Nuvia Ion Exchange Resins post–Protein A capture, to a two-step purification workflow, which includes a Protein A capture step followed by a mixed-mode chromatography step using Nuvia aPrime 4A Resin, a hydrophobic anion exchange resin. We present an additional two-step purification approach using Protein A resin followed by a calcium affinity cation exchange media, CHT Ceramic Hydroxyapatite XT Media. The results of both studies demonstrate that a high purity product can be attained using one fewer unit operation than traditional workflows.

Presented by: William H. Rushton, MS, Process Chromatography Support Scientist and Daniel M. Yoshikawa, PhD, Global Product Manager for Process Chromatography, Bio-Rad Laboratories, Inc.