Continuous Protein A chromatography, such as sequential multi-column chromatography, promises to reduce costs of monoclonal antibody (mAb) capturing steps. Several interconnected columns are loaded at high flow rates, leading to fast processes with high productivities. As a side effect, pressure-flow requirements of continuous processing are challenging. Current rationales for development of continuous processes are based on the adsorption characteristics of the resin. A comparison is drawn between a batch and continuous capturing process for a high capacity TOYOPEARL<sup>®</sup> AF-rProtein A HC-650F resin. Under the conditions tested, the use of continuous processing with a feedstock of 3.4 mg/mL, using 5 x 1 mL prepacked Protein A prototype columns leads to an increase of productivity of over 300%. Additionally, a decrease of buffer and resin usage of over 25% compared to a corresponding batch process using a 5 ml rProtein A SkillPak<sup>®</sup> column and the same feedstock was achieved. The use of 3 columns in the capture zone enabled loading of up to 85% of the SBC of the resin at short residence times of 0.5 minutes, while resulting in a yield of 96.8% and a productivity of 70 g mAb/L resin/h. Residence times of 0.25 minutes were tested as well, resulting in a productivity of 138 g mAb/L resin/h, but resulted in a decreased yield of 86.8 %. Continuous processing requires optimization of the column packing protocol, to counteract the higher flow rates during operation and additional method development for the process transfer. Besides productivity, buffer and resin consumption, product yield and a cost comparison will be presented.

Case study: Process transfer from batch to continuous protein A chromatography

ACS Fall 2021

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