MOESM3 of Isolation of anti-extra-cellular vesicle single-domain antibodies by direct panning on vesicle-enriched fractions

Additional file 3: Figure S3. Antibody differential binding to cell-derived EVs. EVs derived from HEK-293, SKBR3, and Jurkat cells were used to evaluate the binding preferences of the nanobodies H1 and B1 compared with the binding of the irrelevant clone nbVHH (A–C). The binding capacity of a commercial anti-CD9 antibody was tested with the same cell lines (D). Bars indicate median percentage of positively stained EV coated beads with anti-CD9–PE antibodies to H1-GFP, B1-GFP, and non-binding VHH coated beads with respect to autofluorescence of unstained EV coated beads. The error bars indicate standard deviations for triplicate measurements

MOESM4 of Isolation of anti-extra-cellular vesicle single-domain antibodies by direct panning on vesicle-enriched fractions

Additional file 4: Figure S4. Anti-exosome nanobodies bind EV-fractions separated by chromatography. Flow cytometry experiments show that both H1 and H6 strongly bind to exosomes present in the fraction 1 separated by IEX chromatography

MOESM1 of Isolation of anti-extra-cellular vesicle single-domain antibodies by direct panning on vesicle-enriched fractions

Additional file 1: Figure S1. Chromatographic separation of kit-purified EV-enriched fraction from culture media. Kit-precipitated EVs present in SKBR3 cell culture supernatant were separated using a large-pore anion-exchange monolith column. All the four separated fractions were analyzed by flow-cytometry and resulted positive for the EV marker CD9