Open AccessCharacterizing single extracellular vesicles by droplet barcode sequencing for protein analysis
Author(s) -
Banijamali Mahsan,
Höjer Pontus,
Nagy Abel,
Hååg Petra,
Gomero Elizabeth Paz,
Stiller Christiane,
Kaminskyy Vitaliy O.,
Ekman Simon,
Lewensohn Rolf,
Karlström Amelie Eriksson,
Viktorsson Kristina,
Ahmadian Afshin
Publication year - 2022
Publication title -
journal of extracellular vesicles
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.94
H-Index - 68
ISSN - 2001-3078
DOI - 10.1002/jev2.12277
Subject(s) - extracellular vesicles , barcode , vesicle , microvesicles , chemistry , computational biology , extracellular vesicle , extracellular , microbiology and biotechnology , biology , computer science , biochemistry , microrna , membrane , gene , operating system
Small extracellular vesicles (sEVs) have in recent years evolved as a source of biomarkers for disease diagnosis and therapeutic follow up. sEV samples derived from multicellular organisms exhibit a high heterogeneous repertoire of vesicles which current methods based on ensemble measurements cannot capture. In this work we present droplet barcode sequencing for protein analysis (DBS‐Pro) to profile surface proteins on individual sEVs, facilitating identification of sEV‐subtypes within and between samples. The method allows for analysis of multiple proteins through use of DNA barcoded affinity reagents and sequencing as readout. High throughput single vesicle profiling is enabled through compartmentalization of individual sEVs in emulsion droplets followed by droplet barcoding through PCR. In this proof‐of‐concept study we demonstrate that DBS‐Pro allows for analysis of single sEVs, with a mixing rate below 2%. A total of over 120,000 individual sEVs obtained from a NSCLC cell line and from malignant pleural effusion (MPE) fluid of NSCLC patients have been analyzed based on their surface proteins. We also show that the method enables single vesicle surface protein profiling and by extension characterization of sEV‐subtypes, which is essential to identify the cellular origin of vesicles in heterogenous samples.