One of the biotechnology industry's primary challenges is upgrading biomanufacturing processes to align with Industry 4.0. This research focused on microbial fermentation for recombinant protein production, integrating cloud systems, IoT, and AI to enhance process validation. Utilizing the yeast Pichia pastoris and an AI-based respiratory quotient (RQ) control algorithm, significant improvements in protein yields were demonstrated, highlighting the potential of AI-driven physiological control in pharmaceutical and biotech industries' bioprocesses.

To understand the source of interclonal heterogeneity during targeted integration (TI)-based CHO antibody producer cell-line development, we developed a cellular genomic barcoding strategy. This technology provided novel insights about clone diversity during stable cell-line selection on pool level, enabled an imaging-independent monoclonality assessment after single-cell cloning, and eventually improved hit-picking of antibody producer clones by monitoring of cellular lineages during the cell-line development (CLD) process.

Macromolecular complexes are cornerstones of most, if not all, biological processes in cells. We will illustrate how the CRISPR/Cas9 editing technology can be used for gene tagging in order to introduce affinity tags and facilitate the purification of proteins/macromolecular assemblies expressed in physiological conditions. We will also discuss tagging proteins with fluorescent reporters in view of imaging and functional proteomics applications.

We have established and utilised a protein-production pipeline based on cell-free synthesis that can accelerate the production of therapeutic proteins, including cytokines, antibodies, and membrane proteins. Using this pipeline, microgram to milligram quantities of multiple proteins can be produced simultaneously using automated processes, greatly accelerating the production of therapeutic protein candidates for characterisation by analytical methods including NMR. This presentation will highlight the progress of the pipeline.

The combination of global population increase, expected to approach 10 billion by 2050, and rising affluence in developing regions poses a formidable challenge in meeting the predicted 2-fold demand for meat through conventional means. The burgeoning cultivated meat industry presents a promising solution to this sustainability conundrum.

The present work explores an overview of cultivated meat and the following topics:

• Measures to ensure food product safety of the cell lines, cell culture media, processes, and final product testing used to produce cultivated chicken 
• Design specifications, sources, and characteristics of critical nutrients contained in media that impact the cost and mission of cultivated meat
• Bioprocessing at an immense scale compared to traditional biotherapeutics
  

• Critical evaluation of the current industrial evolution
• AI vs mechanistic modelling: what to choose?
• Workflow vs data: where to invest?
• Outlook on how AI will change the way of bioprocessing in the future​

• Definition and examples of hybrid processing
• Benefits for process flexibility and continuous operation
• Technical, logistical and regulatory burdens
• Sustainability considerations 
• Will hybrid processing give a boost for single-use equipment beyond biopharma?​

Cell culture development in biopharmaceuticals uses models to optimise processes and understand complex systems. Defining a model's purpose is crucial, as is starting with good data. Embracing model thinking across all disciplines enhances understanding and effective use of models. Despite challenges, effective modelling can lead to faster, better, and more sustainable processes.

Bio-capacitance has become a standard online method to measure growth in cell-based biomanufacturing. The method offers, rapid continuous monitoring without manual sampling. However, there are noted deviations at the inflection point beyond exponential growth compared to standard staining methods such as trypan blue. This can be explained by different measurement criteria that can be exploited to gain a good understanding of the metabolic changes that arise during the bioprocess.

Digital twins can significantly transform bioprocess design. We present a case study on AAV production in insect cells, where digital simulation was used to design a new fed-batch operation. This demonstrate how digital twins can be leveraged for new gene and cell therapy products, which often have limited accumulated data and serotype/donor-specific variability, to unlock faster and more efficient bioprocess development.

The current workflow involves experimental DOE to determine the optimal culture media formulation. A paradigm shift is underway in the optimisation of culture media, wherein a modelling approach can be employed to accelerate culture media optimisation. I will introduce a computational approach involving genome-scale metabolic modelling and model-guided DoE approach, and how this workflow can help the industry, particularly for new modalities, to accelerate culture media design and optimisation.

Using genome-scale cell-line engineering, we have generated a large collection of engineered CHO cell lines for the production of the next generations of biologics. The collection contains cell lines engineered for prolonged fed-batch productions, removal of lactate secretion, tailored and improved glycosylation, and removal of contaminating host cell proteins. Using these cell lines, we have demonstrated production of recombinant proteins with tailored glycans, lactate-free bioprocesses, and improved product quality.

This presentation will focus on the development and validation of Good Manufacturing Practice-compliant 3D-culture methods using microcarriers and single-use stirred-tank bioreactors for expanding mesenchymal stromal cells (MSCs). Our results showed comparable yields and quality between 3D and 2D cultures, with high cell viability, consistent immunophenotype, and intact multipotency and immunopotency. These processes support scalable, clinical-grade MSC production for use in allogeneic advanced therapies.

Trastuzumab (Herceptin®) is a key treatment for HER2-positive breast cancer. The aim of this study was to develop a stable CHO cell line with high trastuzumab production using a transposon-based vector, suspension culture and eGFP/FACS selection. The T1A7 clone, selected from over 1500 clones, reached 4.24 g/L in 7 days. This efficient workflow provides a cost-effective approach to monoclonal antibody production suitable for R&D laboratories, including those in developing countries.