Integrating Nuclear Magnetic Resonance into Bioprocess Control of Clinical-Grade Extracellular Vesicle Manufacturing

The rapid advance in the research and development of extracellular vesicle (EV)-based therapeutics has stimulated a paradigm shift in the field of regenerative medicine. However, translating EV-based therapies into the clinic requires robust, scalable, and Good Manufacturing Practice (GMP)-compliant bioprocesses that ensure product consistency, potency and safety. In this Perspective, we propose that metabolomics, particularly by using high-resolution nuclear magnetic resonance (NMR), can serve as a transformative analytical technology in EV manufacturing and quality control. By integrating NMR-metabolomics monitoring into upstream cell culture and downstream EV purification workflows, it becomes possible to identify metabolic fingerprints predictive of cell performance, EV yield and EV bioactivity. Drawing from the experience of the GALVANO consortium, which is developing Spain's first GMP-grade platform for EV manufacturing from clinical-grade human Wharton Jelly's mesenchymal stromal cells (WJ-MSC-EVs) with particular promise in the modulation of inflammation and tissue regeneration, we highlight how NMR-metabolomics can support Quality by Design (QbD) principles, enhance in-process analytics and accelerate regulatory harmonisation. We further discuss the need for collaborative standardisation of analytical methods and reporting frameworks to ensure reproducibility and comparability across EV batches. Together, these strategies can advance EV-based therapeutics toward reliable, large-scale clinical application.