Enzyme-linked immunosorbent assay to detect surface marker proteins of extracellular vesicles purified from human urine

The molecular profile of extracellular vesicles released in urine reflects the pathophysiological processes occurring within originating cells located in diverse nephron segments. Here, we present an enzyme-linked immunosorbent assay for quantitative membrane protein detection in extracellular vesicles in human urine samples. We describe steps for preparing urine samples, biotinylated antibodies, and microtiter plates to purify extracellular vesicles and detect membrane-bound biomarkers. The specificity of signals and the limited variability by freeze-thaw cycles or cryopreservation have been verified. For complete details on the use and execution of this protocol, please refer to Takizawa et al. (2022).(1)