This presentation will highlight our manufacturing platform strategy for cell therapies-emphasising data science, modelling, and PAT-to enhance manufacturing efficiency and product quality. By integrating automation and machine learning, we want to accelerate the development of robust processes. Join us in exploring how smart manufacturing practices can redefine the future of cell therapy production.

A novel program of work (Digital_Lyo) will be presented that is being undertaken by a consortium or academic, industrial, and regulatory authority partners, including AstraZeneca, Siemens, the Medicines and Healthcare Regulatory Agency (UK), and smaller industrial enterprises with specialist capability in sensor development. The talk will present highlights of the Digital_Lyo programme, including the applications for a novel process analytical technology called through-vial impedance spectroscopy (TVIS).

For biomanufacturing to reach the standards of Industry 4.0, digital twins are crucial. To accelerate digitalisation efforts in downstream processing, we have developed a framework for automatic generation of mechanistic chromatography models. These models are then used to create a digital shadow of the process by means of Kalman filtering. The result gives improved monitoring of the chromatographic elution, combining both real-time data and mechanistic modelling.

The presented case study investigates the application of automated Raman spectroscopy as an in-line tool for the monitoring of protein aggregation during downstream processing. Spectral effects associated with multiple aggregation phenomena were analysed to provide insights into structural changes of different proteins. Leveraging this knowledge, the potential of Raman spectroscopy for real-time monitoring of critical quality attributes in chromatographic processes was demonstrated.

The production of biopharmaceuticals is both chemical- and water-intensive. The downstream process, which typically involves multiple chromatography steps, requires large volumes of buffers. With the global commitment to sustainable development goals and the anticipated growth of the biopharmaceutical market, buffer consumption is expected to become increasingly problematic. To mitigate this issue, we propose introducing buffer recycling in chromatography to reduce the consumption of water and chemicals.

• ICH Q5 A (R1) Revision: Viral safety Evaluation of Biotechnology 
• Products Derived from Cell Lines of Human or Animal Origin Expectations
• What has changed and the Impact on bioprocessing
• Viral clearance strategies in continuous processing
• Virus removal

• Defining Success Metrics: Key performance indicators for evaluating CMO/CDMO partnerships in bioprocessing projects
• Communication Strategies: Ensuring alignment through clear communication, project updates, and expectations management
• Risk Mitigation: Identifying and addressing common challenges such as quality assurance, timelines, and regulatory compliance
• Scaling and Innovation: Leveraging CMO/CDMO expertise to navigate scale-up/down challenges and integrate emerging technologies

Secreted recombinant proteins often account for a large fraction of the total protein in cultivation supernatants facilitating purification. If, however, cell or tissue disruption is required for product recovery, purification becomes substantially more challenging due to abundant host cell proteins, DNA, and other impurities. Here, we discuss which options exist to build scalable and cost-efficient purification when handling such complex feedstocks.

A model-based strategy is presented for the optimisation of yield and productivity, at a fixed purity specification, in the reversed-phase chromatographic purification of an oligonucleotide. Through this approach, the roles of collection window, column loading, and gradient slope on process performance were elucidated. In addition, it allowed the rapid design of an efficient MCSGP, which, when coupled to a dynamic process controller based on UV signal, ensured a robust operation.

Plasmids are the key starting material for viral vector manufacturing. Due to the rapid growth of the CGT field and increased demand for high-quality pDNA, plasmid supply became a significant bottleneck. Here we present process optimisation using a combination of HTS and mechanistic modelling. The suggested process marks one of the key elements for Roche’s fast growing gene therapy activities, while facilitating the cost decrease of gene therapy products.

Separating bionanoparticles (BNP) from chromatin is particularly challenging due to the wide range of chromatin fragment sizes and charges. While endonuclease treatment reduces host cell DNA, tightly packed DNA within chromatin is resistant to conventional enzymes. Salt-active nucleases effectively remove DNA in chromatin form. This presentation will highlight their use in downstream processing of enveloped bionanoparticles such as HIV-1 gag VLPs and the measles virus.

Toward standardised platform processing-an innovative dual-stage cross-flow filtration setup is introduced for integrated VLP capture and purification. Selective VLP precipitation and washing of the precipitate in the first membrane stage are combined with simultaneous recovery, isolation, and concentration of the re-dissolved VLPs in the second membrane stage. This study spotlights the potential to broaden the setup's applicability to diverse (viral) vectors, distinct process conditions, and continuous processing.

Despite growing interest in extracellular vesicles (EVs), their manufacturing is still limited by low yields and scalability issues. Current separation methods are challenged by co-isolation of impurities, scale-up, and potential changes in physicochemical properties of EVs. Here, we show an isolation strategy for EVs based on zwitterionic polymeric coacervates, exhibiting stimulus-responsiveness, anti-fouling, and liquid-like properties. We demonstrate how this method leads to a versatile, gentle, and scalable purification strategy.

Extracellular vesicles (EVs) facilitate intercellular communication by transporting bioactive molecular cargo and can be exploited as therapeutic agents. However, their purification from physiochemically similar impurities poses a manufacturing challenge. We evaluated the effectiveness of bind-elute ion exchange chromatography (IEX) with a nanofiber-based adsorbent for EV purification. IEX preserved EV bioactivity, and achieved high purity and recovery comparable to widely adopted size-based methods, with the advantage of separating distinct EV subpopulations.