Mimicking continuous capture chromatography for virus clearance using a single chromatography column model.

Continuous chromatography is increasingly being used across the biotechnology industry due to its economic advantages. For adoption in commercial manufacturing, also models for virus clearance studies must be available. It is demonstrated how for a virus clearance study for a multispecific antibody, the continuous protein A capture chromatography process, being run on multiple interconnected columns, can be mimicked with only a single column. With this mimicking small-scale model, resources and complexity can be minimized, when conducting virus clearance studies at a contract research organization (CRO) lab. Obtained log reduction values (LRV) for murine leukemia virus (xMuLV) and minute virus of mice (MVM) virus, used as model viruses, are comparable to batch protein A chromatography and results described by other groups. The feasibility of this mimicking small-scale model helps to further reduce barriers to adoption when implementing continuous chromatography.

Moving to CoPACaPAnA: Implementation of a continuous protein A capture process for antibody applications within an end-to-end single-use GMP manufacturing downstream process.

For the first time to our knowledge the implementation of a continuous protein A capture process for antibody applications (CoPACaPAnA) embedded in an end-to-end single-use 500 L GMP manufacturing downstream process of a multispecific monoclonal antibody (mAb) using a single-use SMB system was conducted. Throughout the last years, a change concerning the pipelines in pharmaceutical industry could be observed, moving to a more heterogeneous portfolio of antibodies, fusion proteins and nanobodies. Trying to adjust purification processes to these new modalities, a higher degree of flexibility and lower operational and capital expenditure is desired. The implementation of single-use equipment is a favored solution for increasing manufacturing agility and it has been demonstrated that continuous processing can be beneficial concerning processing cost and time. Reducing protein A resin resulted in 59% cost reduction for the protein A step, with additional cost reduction also for the intermediate and polishing step due to usage of disposable technology. The downstream process applied here consisted of three chromatography steps that were all conducted on a single-use SMB system, with the capture step being run in continuous mode while intermediate and polishing was conducted in batch mode. Further, two steps dedicated to virus inactivation/ removal and three filtration steps were performed, yielding around 100 g of drug substance going into clinical phase I testing. Therefore, in this study it has been demonstrated that employing a continuous capture within a GMP single-use downstream processing chain is feasible and worthy of consideration among the biotech industry for future application to modality-diverse pipelines.

Scale-up of continuous multicolumn chromatography for the protein a capture step: From bench to clinical manufacturing.

The awareness about implementing continuous processing for biopharmaceutical products has significantly increased throughout the recent years not only at developmental scale but also for phase I supply in clinical trial manufacturing. In this study, we focused on upscaling continuous protein A chromatography from lab to pilot scale using the Cadence™ BioSMB PD and the Cadence™ BioSMB Process 80 system, respectively. Additionally, we evaluated hardware and software capability whilst running the system for 10 days non-stop using feed from a perfusion bioreactor. In terms of product quality and removal of impurities, comparable data was obtained regarding lab scale and production scale. Compared to batch mode, productivity was increased by 400 to 500%. Furthermore, the system worked accurately during the whole trial, proving its potential for the implementation into a hybrid or an end-to-end continuous process.

Feasibility of using continuous chromatography in downstream processing: Comparison of costs and product quality for a hybrid process vs. a conventional batch process.

The protein A capture step is the main cost-driver in downstream processing, with high attrition costs especially when using protein A resin not until end of resin lifetime. Here we describe a feasibility study, transferring a batch downstream process to a hybrid process, aimed at replacing batch protein A capture chromatography with a continuous capture step, while leaving the polishing steps unchanged to minimize required process adaptations compared to a batch process. 35g of antibody were purified using the hybrid approach, resulting in comparable product quality and step yield compared to the batch process. Productivity for the protein A step could be increased up to 420%, reducing buffer amounts by 30-40% and showing robustness for at least 48h continuous run time. Additionally, to enable its potential application in a clinical trial manufacturing environment cost of goods were compared for the protein A step between hybrid process and batch process, showing a 300% cost reduction, depending on processed volumes and batch cycles.