The ISEV workshop “Extracellular Vesicle (EV)-based biomarkers” aims to discuss key challenges in commercialisation and clinical implementation of EV-based biomarkers. The biomarker potential of EVs, and their application as liquid biopsies, was first demonstrated over two decades ago. Whilst considerable interest in using EVs as biomarkers for a wide range of diseases exists, progress in the field is hindered by a lack of development of EV-based biomarkers under formal regulatory requirements. For example, The Therapeutic Goods Administration (TGA, like the FDA in the USA) in Australia clearly defines the pathway for the clinical implementation of all new diagnostic tests and the clinical evidence appropriate for its intended use and risk classification (Therapeutic Goods (Medical Devices) Regulations 2002). Specific requirements that must be satisfied to ensure a feasible pathway for clinical implementation and compliance with the conformity assessment procedure of the Medical Devices Regulations include evidencing, analytical performance, clinical performance, and clinical utility. Thus, this workshop aims to provide a platform to discuss challenges of the EV community in translating EV-based biomarkers into clinical practice, with focus on the regulatory requirements and commercial aspects.