Profiling surface proteins on individual exosomes using a proximity barcoding assay
2019
Exosomeshave been implicated in numerous biological processes, and they may serve as important disease markers. Surface proteins on
exosomescarry information about their tissues of origin. Because of the heterogeneity of
exosomesit is desirable to investigate them individually, but this has so far remained impractical. Here, we demonstrate a proximity-dependent
barcodingassay to profile surface proteins of individual
exosomesusing antibody-DNA conjugates and next-generation sequencing. We first validate the method using artificial
streptavidin-oligonucleotide complexes, followed by analysis of the variable composition of surface proteins on individual
exosomes, derived from human body fluids or
cell culture media.
Exosomesfrom different sources are characterized by the presence of specific combinations of surface proteins and their abundance, allowing
exosomesto be separately quantified in mixed samples to serve as markers for tissue-specific engagement in disease. The use of antibodies to capture and profile
exosomeslimits the number of target proteins that can be detected. Here the authors develop a proximity-dependent
barcodingassay that allows profiling of 38 surface proteins on individual
exosomesfrom heterogeneous samples such as serum and seminal fluid.
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