One-step integrated clarification and purification of a monoclonal antibody using Protein A Mag Sepharose beads and a cGMP-compliant high-gradient magnetic separator

• Published in: New biotechnology Vol. 42; pp. 48 - 55
• Main Authors: Ebeler, Moritz, Lind, Ola, Norrman, Nils, Palmgren, Ronnie, Franzreb, Matthias
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 25.05.2018; Elsevier Science Ltd
• Subjects: Beads; Biopharmaceuticals; Biotechnology; Cell culture; Cyclic GMP; Direct capturing; GMP; High-gradient magnetic separation; Industrial biotechnology; mAb downstream processing; Magnetic separation; Monoclonal antibodies; Protein A; Protein purification; Proteins; Purification; mAb downstream processing; GMP; Industrial biotechnology; Direct capturing; High-gradient magnetic separation
• ISSN: 1871-6784; 1876-4347
• DOI: 10.1016/j.nbt.2018.02.007

[Display omitted] •Direct capturing of a mAb from CHO cell culture by high-gradient magnetic separation.•Technical scale process development for a commercial GMP-compliant HGMS device.•In situ product removal using protein A magnetic particles without cell damage. Monoclonal antibodies are a dominant component of today's biopharmaceutical market and are typically purified by classical platform processes. However, high costs and rising demands are drivers for the development of new, efficient and flexible integrated purification processes. Currently, high-gradient magnetic separation as a direct capturing tool for protein purification suffers from the lack of suitable GMP-compliant separation equipment for industrial scale. As a solution for this bottleneck, we present a purification process for a monoclonal antibody directly from CHO cell culture by use of protein A-functionalized magnetic particles together with the first pilot-scale GMP-compliant ‘rotor-stator’ high-gradient magnetic separator. Five consecutive purification cycles were performed, achieving consistent yields of over 85% and purities of over 95%. Stable cell viabilities during the magnetic separation process enable integration of the device as an in situ product removal tool. A comparison with state-of-the-art protein A column-based purification processes reveals a 3-times higher process productivity per mL of applied resin and demonstrates the great potential of magnetic separation in downstream processing.