Process considerations for Protein A affinity capture, virus inactivation, and linked polishing steps in multi-column continuous purification of monoclonal antibodies

摘要

1. Typical mAb purification platform catch chromatography slept have been converted to continuous multi-column chromatography using Semba ProComposerTM software and then quickly and easily tesled with the ProPD™ System. 2. Protein A capture productivities of 80-112 g/L/h were obtained using three different CHO culture fluid feed streams with mAb titers of 7-10.8 g/L and eight 5-ml columns. 3. Scaled comparison of a typical 2.000 L 5 g.'L mAb titer Protem A batch capture veraus a continuous 8-column process showed significant reduction in consumables and overall processing time. Protein A resin volume was decreased 7.1-fold buffer volume decreased 2.9-fold process time decreased from 7 days to 1 day and process productivity increased 14-fold. 4. Polishing by continuous 2-column AEX and 6-column Mixed Mode required only three buffers 'or both processes, and reduced aggregate to 1.5%, HCP by 2.6 logs DNA to 19 pg/mg, and residual Protein A to < 0.1 ppm. 5. Step yields for the continuous Protein A capture. AEX. and MM processes were all > 90% with a final overall yield of 77% .