The empty and full capsid separation for recombinant adeno-associated virus (rAAV) products is a hot topic in rAAV gene therapy community. By adopting weak portioning chromatography (WPC), empty-AAV could be displaced by full-AAV during overloading of the AEX column, resulting in improved full-AAV percentage in AEX eluate. In addition, by harnessing the power of multi-column chromatography (MCC) technology, the cost of goods (COGs) reduction was further decreased. Compared with normal batch AEX operation, AEX-WPC-MCC operation demonstrated its superiority in both full-AAV percentage and gene of interest (GOI) recovery.

Continuous cell expansion of adherent cell lines has the potential to save considerable time and money if it can successfully be integrated into bioprocessing. We have demonstrated that a single N-1 reactor growing anchorage dependent cells on microcarriers is able to run for weeks, supplying adequate cell mass for multiple iterations of an LVV production process. This has the potential to accelerate processing time, allowing manufacturing facilities to downsize their footprint, scale back equipment requirements, and limit workforce demand.

With large potential demand for antisense oligonucleotides on the horizon, significant increases in downstream throughput are becoming extremely important. Continuous chromatography approaches, including CaptureSMB and Multicolumn Countercurrent Solvent Gradient Purification (MCSGP), can provide the throughput needed, while also maintaining desired purity. Utilizing two columns in a continuous mode for aqueous purification of antisense oligonucleotides demonstrated a near four-fold increase in productivity for the HIC step of a model process.

The status quo in operational software for manufacturing is not designed to enable continuous manufacturing. Capturing data for the entire product lifecycle, building a data-based AI model and/or digital twin, capturing inline monitoring data, and then being able to quickly compute process health requires multiple, expensive, legacy software packages that do not inherently interoperate.  Continuous biomanufacturing requires integrated software that leverages the latest technologies to do all the above; to quickly and accurately assess past, current, and future states; and to provide prescriptive recommendations or direct control to keep biomanufacturing processes running optimally.     

The rapid growth and development of new therapeutic modalities from large molecules to cellular therapies have intensified pressure to increase processing throughput while decreasing cost and turnaround time. Multi-omic technologies including genes and metabolomics are critical to understanding the biological basis of new process development. Metabolomics is unique in providing systems-wide small molecule-derived biochemical insights for unbiased data-driven approaches for bioprocessing, from initial designs to scaling-associated quality. Optimizing bioproduction across the product development lifecycle using metabolomics will be discussed with specific case studies on optimization of cell bioprocess, data-driven DoE optimization, and integration with multi-omic technology for strain engineering. 

Continuous processing is the holy grail for many industries and became popular for bioprocessing in the last decade, too. Intensification is a prerequisite to enable a step-wise transformation toward that goal. This presentation gives a comprehensive overview of strategies where and how to implement process intensification, quantifies the benefits like plant occupancy time, and optimizing capacity based on successful examples and case studies.

With the move to intensive processing there is a strong desire to understand the holistic impact of the manufacturing operation on the environment and the business efficiency. The latest process models evaluate facility efficiency (doses per unit volume of cleanroom), PMI, and total energy efficiency. Pre-release versions were used by Process Intensification team in NIIMBL to support sustainability assessments. By way of example, comparisons are made between standard fed-batch processes and intensified process options that include perfusion and continuous downstream operations at different scales. 

Continuous biomanufacturing can intensify existing batch processes, potentially achieving greater efficiencies with a smaller footprint. However, successful execution of continuous biomanufacturing can be more complex than traditional batch processes because it requires maintaining a steady-state operation. Thus, continuous biomanufacturing processes may benefit from robust in-line PAT for real-time monitoring and control of culture state and product quality. This talk will explore current PAT for use in continuous bioreactor cultures.

Continuous Manufacturing (CM) strategies for the production of pharmaceutical drug substance and drug product are receiving increasing attention from manufacturing companies. Several small molecule products for which at least some CM steps have been implemented have recently received approval by regulatory authorities. Implementation of CM for biologics has lagged behind small molecules. This presentation will discuss the opportunities for CM in large molecule manufacturing, with a focus on the critical role that PAT implementation will play for these processes.

Recent approaches in the biopharmaceutical industry are focusing on process intensification and digital transformation. Some of these new capabilities have the potential to significantly change process development. A showcase for a continuous/connected process and the use of a predictive control architecture utilizing digital twins will be presented. Some specific early benefits for process development, optimization, and control will be highlighted – as well as a forecast of future directions.

Interest in continuous biomanufacturing has recently increased due to the potential for a more efficient process with economical advantages. However, there are new challenges to be solved before these processes are implemented in GMP environments. The implementation of PAT, especially with single-use sensors, is critical for continuous bioprocessing. This presentation will review the requirements for utilizing single-use sensors in a continuous process using PendoTECH sensors as a case study for qualification. 

We will present the development activities associated with the selection of a high-capacity Protein A resin for a biologics continuous manufacturing process. The work herein discusses the results of bench scale studies including resin screening of various high-capacity and base stable resins, followed by process optimization with considerations for a continuous manufacturing application. We will include small scale and pilot scale data generated for resin lifetime under different regeneration conditions, as well as impact of bioburden control strategy on long term resin stability. The optimal conditions from bench and pilot runs were implemented at a 500L scale using a continuous manufacturing process.

The commercialized batch-mode process is converted into a continuous MCC process on the laboratory scale. The steps of the MCC development, the required data packages, and the challenges and technical difficulties are illustrated in this presentation.

N-1 perfusion is one strategy for intensifying fed-batch processes. This talk will focus on the intensification of a fed-batch process using an ambr250HT to run N-1 perfusion cultures and inoculate fed-batch vessels from them.

Accessibility and affordability of biologics globally is poor. Significant changes in development, manufacturing, and distribution are required for the broad use of life-saving biologics in low- and middle-income countries. Disruptive innovations that could improve global access, such as small footprint biomanufacturing, are enabled by integrated, intensified, and automated processes. Here, we discuss the development and optimization of straight-through purification processes and demonstrate their utility in small footprint, automated, end-to-end manufacturing.

Process intensification has already been demonstrated as a tool for increasing the productivity of monoclonals. Nevertheless, several challenges remain in what concerns virus manufacturing. In this talk, we will highlight perfusion, periodic counter-current chromatography, and continuous filtration operations as enabling tools for a streamlined AAV manufacturing process, focusing on the caveats and pitfalls of the process development journey.

The upstream process development team at Pfizer has developed a highly productive continuous bioreactor process. However, with the implementation of perfusion, traditional bench-scale models become highly labor intensive with a lowered throughput that makes practical process development and characterization challenging. An automated and high-throughput solution for efficient development of an intensified continuous process in the ambr250 HT perfusion system is presented.

This work demonstrates a showcase of a truly continuous and automated process for the production and purification of a monoclonal antibody. It highlights, both, the potential of using protein precipitation as a capture step, and the issues and challenges in designing and constructing of such a process skid.