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Published on: September 12, 2019
Protocol for isolating human extracellular vesicles from nephrotic urine and enriching autoimmunoglobulin-triggered
Karen Lahme1, Catherine Meyer-Schwesinger1
1Institute of Cellular and Integrative Physiology, Center for Experimental Medicine, University Medical Center Hamburg-Eppendorf (UKE), 20246 Hamburg, Germany; Hamburg Center of Kidney Health, University Medical Center Hamburg-Eppendorf (UKE), 20246 Hamburg, Germany.
This protocol describes the isolation of total urinary extracellular vesicles (EVs) from nephrotic patient urines and enrichment of AutoImmunoglobulin-Triggered Extracellular Vesicles (AIT-EVs). Procedures for urine collection, sample processing, buffer preparation, antibody coupling to DynaBeads, urine ultrafiltration, and total EV isolation are provided. The protocol further details EV concentration measurement and selective AIT-EV capture and elution using anti-human IgG4 antibodies. This approach enables reproducible enrichment of AIT-EVs and can be adapted to isolate other EV subtypes by substituting the capture antibody.
This protocol describes the isolation of total urinary extracellular vesicles (EVs) from nephrotic patient urines and enrichment of AutoImmunoglobulin-Triggered Extracellular Vesicles (AIT-EVs). Procedures for urine collection, sample processing, buffer preparation, antibody coupling to DynaBeads, urine ultrafiltration, and total EV isolation are provided. The protocol further details EV concentration measurement and selective AIT-EV capture and elution using anti-human IgG4 antibodies. This approach enables reproducible enrichment of AIT-EVs and can be adapted to isolate other EV subtypes by substituting the capture antibody.
