Profiling surface proteins on individual exosomes using a proximity barcoding assay.

Wu, Di; Yan, Junhong; Shen, Xia; Sun, Yu; Thulin, Måns; Cai, Yanling; Wik, Lotta; Shen, Qiujin; Oelrich, Johan; Qian, Xiaoyan; Dubois, K. Louise; Ronquist, K. Göran; Nilsson, Mats; Landegren, Ulf; Kamali-Moghaddam, Masood

Exosomes have been implicated in numerous biological processes, and they may serve as important disease markers. Surface proteins on exosomes carry information about their tissues of origin. Because of the heterogeneity of exosomes it is desirable to investigate them individually, but this has so far remained impractical. Here, we demonstrate a proximity-dependent barcoding assay to profile surface proteins of individual exosomes using 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. Exosomes from different sources are characterized by the presence of specific combinations of surface proteins and their abundance, allowing exosomes to be separately quantified in mixed samples to serve as markers for tissue-specific engagement in disease. The use of antibodies to capture and profile exosomes limits the number of target proteins that can be detected. Here the authors develop a proximity-dependent barcoding assay that allows profiling of 38 surface proteins on individual exosomes from heterogeneous samples such as serum and seminal fluid.

EXOSOMES; BODY fluids; STREPTAVIDIN; HUMAN body; PROTEINS; NUCLEOTIDE sequencing; CELL culture; OLIGONUCLEOTIDES

Nature Communications, 2019, Vol 10, Issue 1, pN.PAG

2041-1723

Academic Journal

10.1038/s41467-019-11486-1