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Pawp (Protein A Well-plate Purification) a High Throughput Method for Purification of Monoclonal Antibodies

Jennifer Hopp, Ross Pritchett, Maria Darlucio, Junfen Ma, and Judy H. Chou. Process Research & Development, Genentech, Inc., One Antibody Way, Oceanside, CA 92056

High throughput techniques for purification of monoclonal antibodies from harvested cell culture fluid can dramatically minimize feedstock requirements and accelerate process development decision-making during clone selection and cell culture process optimization. In this presentation, development of a scale-down high throughput antibody purification technique will be described. This method uses Protein A resin in a 96 well-plate format yielding sufficient quantity and concentration of antibody materials to perform multiple analyses of product quality per well. Protein A resins were selected and buffer conditions were optimized to obtain aggregate and charge variance comparability with antibodies purified by preparative Protein A chromatography. This comparability was demonstrated with three model monoclonal antibodies by size-exclusion chromatography, ion-exchange chromatography, imaged capillary isoelectric focusing, capillary zone electrophoresis, and glycan identification by capillary electrophoresis. The developed method can be performed in manual format and robotic format. It also maintains flexibility of resin types and amounts and offers cost-savings over commercially available products.