The transition from traditional fed-batch to high-density perfusion strategies is a critical strategy for enhancing productivity in biopharmaceutical manufacturing. Using the NISTCHO, a standardized Chinese Hamster Ovary (CHO) platform producing the cNISTmAb monoclonal antibody, reference cell line, we demonstrate how perfusion-based processes can significantly reduce costs while maximizing product yield. We evaluated NISTCHO in a simulated, long-term perfusion culture. Main output was achieving high viability and sustaining very high cell densities, which resulted in a higher volumetric productivity compared to standard fed-batch systems. By continuously replacing media and removing inhibitory by-products, the system maintains superior culture health, facilitating a 2-fold higher specific productivity. Furthermore, implementing N-1 perfusion, where the NISTCHO seed train is cultured in a high-density perfusion environment, allows for inoculation of the production bioreactor at 5–10 fold higher densities, substantially reducing the production phase duration. This "intensification" reduces facility footprint and lowers operational expenses (media usage, labor) while ensuring consistent product quality. These results provide a robust, scalable framework using the NISTCHO standard, demonstrating that transitioning to perfusion-based, continuous production, especially at the N-1 step, is a powerful approach for optimizing the balance between productivity and cost-effectiveness in modern biologics manufacturing.

This talk explores how targeted media design enables intensified bioprocessing by supporting higher cell densities, sustained productivity, and improved product quality. It will highlight strategies for optimizing nutrient composition, feeding approaches to enhance manufacturing efficiency in intensified bioprocessing systems.

This presentation explores how different feeding strategies influence cellular behavior and performance in high-intensity dynamic perfusion bioprocesses.

The perfusion culture of mammalian cells producing biopharmaceuticals is essential for integrative continuous bioprocessing, which has been explored extensively for benefits in biomanufacturing. For several decades, various perfusion culture systems using cell retention devices such as spin filter, centrifuge, sonic decanter, and membrane filters have been introduced successfully only for short-term cultures. For seamless integrative continuous bioprocessing, connecting the upstream cell culture processes to the downstream purification processes, ‘the long-term stability’ of perfusion cultures needs to be strongly secured. In this presentation, author’s research experiences on perfusion culture systems and efforts for stable long-term perfusion cultures will be shared.

This talk explores strategies to optimize tangential flow filtration (TFF)–based perfusion systems to support efficient continuous biomanufacturing. It will highlight approaches to improve process productivity and streamline operations while reducing equipment demands and facility footprint.

In a bid to sustainably meet the growing need for mAbs, we have developed a fully continuous, precipitation-based downstream process, drawing inspiration from the plasma fractionation industry. This new downstream process can be greater in capacity, less raw material-intensive, and significantly cheaper than the platform process. We’ll describe the genesis, evolution, and optimization of the process in terms of mAb critical quality attributes and sustainability metrics, and the path forward.

Continuous bioprocessing offers transformative potential but faces persistent scale-down and integration challenges. In downstream processing, maintaining stable micro- to milliliter-per-minute flow rates is essential, and fully continuous solid–liquid separation remains a bottleneck. Hollow-fiber based two-stage filtration provides a practical scale-down strategy, while continuous precipitation and flocculation represent powerful alternatives when robust separation is ensured. At manufacturing scale, fluid and buffer logistics dominate. Emerging soft-sensor concepts for online CQAs enable long-running, integrated processes, positioning continuous biomanufacturing as a cornerstone for future carbon-neutral pharmaceutical production.

Antibody conjugation through mAb-engineered cysteine sites is a method to achieve drug-to-antibody (DAR) selectivity for ADCs. This talk describes utilization of process analytical tools (PAT) to develop a continuous, ultrafiltration decapping reaction. HP-IEX monitored real time decapping of mixed thiol caps. A redox probe correlated redox potential to decapping efficiency. Continuous decapping was scaled to generate multi-kilogram quantities of decapped mAb, which resulted in high-purity ADC.

Plasmid DNA (pDNA) underpins viral vectors, mRNA vaccines, and DNA therapeutics, yet production is limited by costly, low-yield batch processes. iCAP transforms this with a continuous, automated microbial platform integrating bioprocess intensification and digital intelligence. Using genetically stabilized E. coli and a growth-decoupled replication system, iCAP enables stable, long-term plasmid production in bioreactor cascades. Continuous alkaline lysis, chromatographic purification, and real-time digital-twin–based control ensure high-throughput, low-variability recovery. Modular and scalable, iCAP reduces costs and environmental impact while supporting decentralized vaccine manufacturing and integrated therapeutic supply, setting a new benchmark for sustainable, next-generation plasmid DNA biomanufacturing.

This presentation will compare the production of Fibroblast Growth Factor-2 (FGF2) using two microbial hosts with distinct bioprocess characteristics, necessitating different strategies for process optimization. This talk will provide the audience with a practical framework for selecting and optimizing microbial hosts for growth factor production based on their intrinsic biological and bioprocess characteristics. The presentation will also demonstrate how kinetic modeling can be directly translated into operational decisions for continuous and perfusion cultures, and how advanced fermentation strategies such as chemostat-filtration coupling and cascade fermentation can be used to overcome metabolic burden and stability limitations. These concepts will be broadly applicable to the continuous manufacturing of recombinant proteins for biopharmaceutical, cultivated meat, and serum-free media applications.

A 785 nm Raman-based model was developed as an online measurement for the detergent concentration in a continuous viral inactivation (VI) step. In the VI step, detergent is continuously added to a concentrated bioreactor perfusate stream. The Raman flow cell was first calibrated using a bank of previously generated perfusate samples spiked to varying detergent levels to generate a PLS model with an estimated accuracy of 0.02% w/v.

This talk discusses the world’s first digital twin for continuous lyophilization of biotherapeutics in suspended vials. This digital twin consists of a state-of-the-art mechanistic model, a state observer for real-time tracking of the residual moisture, a highly efficient framework for uncertainty analysis, and a novel dynamic optimization algorithm. Combining all components, this virtual representation of the continuous lyophilization system can be used for improving the manufacturing in real time.

A digital shadow is a near–real-time, data-driven virtual representation of a bioreactor and its process. An AI-augmented digital shadow (AIS) accelerates process intensification by integrating multivariate data with machine-learning and hybrid mechanistic models to identify actionable parameters and nonlinear interactions. AIS enables what-if simulation of equipment configurations, CPP strategies, and feeding profiles before execution. It can infer hard-to-measure state variables, assess scale equivalence, and support model-predictive and adaptive control by recommending near-optimal setpoints. By mapping high-dimensional parameter space and ranking limiting phenomena, AIS strengthens design-space definition, regulatory justification, first-time-right success, and manufacturing robustness.