In this presentation, we will address major challenges in rAAV-based gene therapy manufacturing, highlighting two recent breakthroughs in the HeLaS3 production platform: (1) a novel cell engineering strategy to boost productivity under intensified conditions, achieving up to 10-fold higher yields, and (2) a streamlined selection system that accelerates stable cell line generation to ~2 months while maintaining high titers and vector quality. Together, these approaches tackle key bottlenecks in rAAV production.

Despite their central role in biopharma, HEK293 cells are genomically unstable, making reproducibility a persistent challenge. Whole-genome datasets from multiple laboratories were analysed to construct a HEK293-specific, viral-augmented reference. Compared with the standard human genome, this reference improves mapping fidelity, stabilises copy-number detection, and reduces artefactual variation. When combined with cohort-based thresholds, these advances create a genomic QC framework that enables consistent characterisation of HEK293 derivatives and strengthens confidence in cell line-based bioprocess development.

Macromolecular complexes are fundamental to nearly all cellular biological processes. This presentation will illustrate how CRISPR/Cas9 genome editing can be applied for gene tagging, enabling the insertion of affinity tags to simplify the purification of proteins and macromolecular assemblies produced under physiological conditions. The use of fluorescent reporter tags for proteins, highlighting their applications in imaging and functional proteomics will also be discussed.

This talk introduces a multiplexable orthogonal transposon system for advanced bioprocessing. We demonstrate its use in CHO cells for both gene knock-in and knockdown, allowing for precise control of cellular physiology. This tool enables the creation of new host cell lines and the modulation of product quality. Our findings are supported by data confirming precise integration and long-term stability, showcasing a significant step forward in engineering next-generation biotherapeutic cell lines.

In this presentation, we will explore how single-cell omics technologies are advancing our understanding of biological systems, with a focus on monoclonal antibody–producing CHO cell lines. Our studies integrate measurements of chromatin accessibility, gene expression, and protein abundance at single-cell resolution to uncover cellular heterogeneity and regulatory mechanisms. We will also highlight the platforms used for single-cell profiling and the computational strategies employed to analyse and interpret these data.

Although Integrated Continuous Biomanufacturing can greatly improve recombinant protein space-time yields, fed-batch remains dominant because it limits the emergence of undesirable subpopulations. Here, we demonstrate how this metabolic behavior is controlled using cascaded continuous processing. The trajectory of specific productivity is described as a function of accumulated cell effort, basis for smooth control via multiple feeds. The process was scaled to pilot scale using automation and digital twins.

The demand for recombinant adeno-associated virus (rAAV) vectors is driving the need for scalable, efficient manufacturing solutions. Herein we describe a novel AAV perfusion-enhanced expression (APEX) process, resulting in a 3–6 fold increase in volumetric productivity while maintaining robust and consistent product quality in Pinnacle PCLTM cell lines. This process has been successfully demonstrated across multiple serotypes in large scale cell culture with titers approaching 1 × 1012 GC/mL. This presentation will highlight the development and implementation of the APEX perfusion strategy from bench to pilot-scale 250L bioreactors and discuss scale-up challenges, including: identifying optimal perfusion rates to attenuate metabolic waste production, reconciling media preparation concerns, addressing bioreactor challenges with increased cell mass, and additional intensification of harvest clarification to enable forward processing with higher levels of cell debris and impurities. The APEX production platform marks a significant leap forward in the efficient and scalable manufacturing of rAAV vector products.

Bioprocess control involves managing non-linear, time-evolving cell populations, unlike traditional chemical or pharmaceutical processes. This complexity requires adaptive strategies supported by multivariate monitoring and historical data to inform control actions. In this work, we propose a novel data-driven control scheme for fed-batch upstream operations, using graph theory to identify dynamic control parameters beyond conventional ones. Machine learning and optimisation enable real-time recalibration and reactive control based on historical insights. This closed-loop, multi-attribute model ensures cultures follow a desired trajectory, effectively addressing variability and enhancing control across diverse conditions, cell types, and products where static strategies may fall short.

During the course of a bioprocess strain, physiology is likely to change. By natural selection, the performance of the cultivation can be positively affected—or more often—during the production of recombinant proteins the culture is degenerated and loses its metabolic capabilities. Under usage of models this changes can be foreseen and preventative control actions can occur to stabilize the cultivation. It will be shown how to measure in real-time adaption and degeneration in E. coli and C. glutamicum cultivations and how to use this information to optimise and stabilise the process?

Continuous bioprocesses on a small scale are becoming increasingly important due to rising yields, improved product quality, and the shift toward personalised medicine. This presentation introduces 3D-printed microfluidic systems for efficient cell retention in perfusion processes. It also features a miniaturised periodic counter-current chromatography system for continuous monoclonal antibody purification, providing a compact, integrated solution for streamlined upstream and downstream bioprocessing.

Cell and gene therapy manufacturing requires seamless integration across upstream and downstream processes to ensure efficiency, consistency, and regulatory compliance. This presentation will explore strategies for unifying process data, enabling end-to-end visibility and control. Case studies will demonstrate how integrated data platforms and advanced analytics improve decision-making, support technology transfer, and enhance scalability—helping organisations accelerate development and deliver high-quality therapies more reliably to patients.

Chinese hamster ovary (CHO) cells remain the dominant host for biologics production, yet metabolic imbalances can generate toxic byproducts that limit cell growth and product quality. This presentation will discuss engineering strategies to reprogram CHO metabolism and reduce accumulation of inhibitory compounds. Examples will highlight genetic modifications, pathway optimisation, and analytical approaches that enable healthier cultures, improved productivity, and more consistent performance in large-scale biomanufacturing.