Open AccessSingle Extracellular Vesicle Protein Analysis Using Immuno‐Droplet Digital Polymerase Chain Reaction Amplification
Author(s) -
Ko Jina,
Wang Yongcheng,
Carlson Jonathan C. T.,
Marquard Angela,
Gungabeesoon Jeremy,
Charest Alain,
Weitz David,
Pittet Mikael J.,
Weissleder Ralph
Publication year - 2020
Publication title -
advanced biosystems
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.153
H-Index - 18
ISSN - 2366-7478
DOI - 10.1002/adbi.201900307
Subject(s) - extracellular vesicles , multiplex , digital polymerase chain reaction , microfluidics , chemistry , biophysics , vesicle , extracellular vesicle , polymerase chain reaction , polymerase , fluorescence , dna , nanotechnology , biological system , microbiology and biotechnology , computational biology , materials science , microvesicles , biology , biochemistry , physics , gene , bioinformatics , microrna , membrane , quantum mechanics
There is a need for novel analytical techniques to study the composition of single extracellular vesicles (EV). Such techniques are required to improve the understanding of heterogeneous EV populations, to allow identification of unique subpopulations, and to enable earlier and more sensitive disease detection. Because of the small size of EV and their low protein content, ultrahigh sensitivity technologies are required. Here, an immuno‐droplet digital polymerase chain reaction (iddPCR) amplification method is described that allows multiplexed single EV protein profiling. Antibody–DNA conjugates are used to label EV, followed by stochastic microfluidic incorporation of single EV into droplets. In situ PCR with fluorescent reporter probes converts and amplifies the barcode signal for subsequent read‐out by droplet imaging. In these proof‐of‐principle studies, it is shown that multiplex protein analysis is possible in single EV, opening the door for future analyses.