The scientific community is currently experiencing an outpouring of research surrounding extracellular vesicles (EVs) and, more specifically, exosomes. This is due not only to their critical role in intercellular communication, but also to their potential to be used as diagnostic tools and/or therapeutic agents in a wide range of pathological conditions1. The rising interest in exosomes, coupled with the immense volume of research, underlies significant needs for both the isolation of high quality exosomes from large-scale batches and the development of industry standards for the characterization and quality control testing of exosomes. While traditional methods, which include the use of ultra-centrifugation and density gradients, are suitable for small-scale studies, the development of scalable and robust processes for the isolation of functional exosomes is essential to meet the growing needs of the scientific community. Here, we report the use of tangential flow filtration (TFF) for the isolation and concentration of functional exosomes from large batches of conditioned culture medium to include lung carcinoma cells, human mesenchymal stem cells (MSCs), and human induced pluripotent stem cells (iPSCs).