Presentations include:
Process considerations for development of multi-column continuous capture of monoclonal antibodies
Anthony Grabski, Ph.D., Director of R&D, Semba Biosciences, Inc., Madison, WI
Multi-column continuous chromatography (MCC) offers significant economic advantages over traditional batch methods for purification of monoclonal antibodies (mAbs), including increased resin capacity utilization, smaller columns, reduced buffer consumption, and faster process time. The Protein A capture step is a primary target to apply MCC due to its high cost, which is driven even higher as improvements in upstream processing have produced a steady increase in mAb titers. In this study we consider the key factors in designing an MCC process that optimizes productivity. Process design begins with determination of the feed duration, which is governed by the mAb titer, column dimensions, residence time, binding capacity, and flow properties of the resin. Use of 3 columns in the capture zone enables efficient utilization of the resin even at short residence times, thereby increasing productivity. Column number and resin volume are optimized to relieve scheduling constraints experienced at high mAb titer while maximizing cycling for single use columns.
Productivities exceeding 100 g mAb/L resin/h were achieved using the Semba ProPD™ System with a concentrated CHO cell culture fluid and 8 x 5 ml TOYOPEARL® AF-rProtein A HC-650F columns. We scaled up to process 100-grams of mAb and directly compared an optimized single-column batch process to an MCC process using two different Protein A affinity resins. A 2.46-liter column was used for the batch runs and eight 50-ml columns were used for the MCC runs on the Semba ProPD System. Resins used for this study included TOYOPEARL AF-rProtein A HC-650F (Tosoh Bioscience) and MabSelect SuRe™ (GE Healthcare). In each run, CHO cell culture fluid containing approximately 100 grams of mAb harvested from the same 250 liter single-use bioreactor run were processed using the identical buffer platform and seven processing steps. Results including process yield, product quality, buffer utilization, and process productivities will be presented.
Scalable pre-packed column performance: 1 mL high throughput screening columns to 57 L production scale columns
Adam Nelson, Ph.D., OPUS Technical Specialist and Fletcher Malcom, MBA, Director of Product Management, Repligen Corporation, Waltham, MA
Pre-packed chromatography columns are routinely used in downstream process development and scale-down studies. In recent years they have also been widely adopted for large scale cGMP manufacturing of biopharmaceuticals. Despite widespread adoption, consistent chromatographic performance data for pre-packed columns used in protein separations at different scales has not yet been demonstrated. In our case studies, we demonstrate performance results obtained with small scale columns (0.5 cm diameter x 5 cm length, 1 mL column volume) that are scalable to production sized columns (60 cm diameter x 20 cm length, 57 L column volume).
