Multiplex protein profiling method for extracellular vesicle protein detection

Abstract Extracellular vesicles (EVs) are small nanometer-sized membrane sacs secreted into biological fluids by all cells. EVs encapsulate proteins, RNAs and metabolites from its origin cell and play important roles in intercellular communication events. Over the past decade, EVs have become a new...

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Autores principales:	Li Sun, David G. Meckes
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9134c49c675a4c8c9f16183c8492ad6a
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spelling	oai:doaj.org-article:9134c49c675a4c8c9f16183c8492ad6a2021-12-02T17:41:31ZMultiplex protein profiling method for extracellular vesicle protein detection10.1038/s41598-021-92012-62045-2322https://doaj.org/article/9134c49c675a4c8c9f16183c8492ad6a2021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-92012-6https://doaj.org/toc/2045-2322Abstract Extracellular vesicles (EVs) are small nanometer-sized membrane sacs secreted into biological fluids by all cells. EVs encapsulate proteins, RNAs and metabolites from its origin cell and play important roles in intercellular communication events. Over the past decade, EVs have become a new emerging source for cancer diagnostics. One of the challenges in the study of EVs and there utility as diagnostic biomarkers is the amount of EVs needed for traditional protein analysis methods. Here, we present a new immuno-PCR method that takes advantage of commercially available TotalSeq antibodies containing DNA conjugated oligos to identify immobilized protein analysts using real-time qPCR. Using this method, we demonstrate that multiple EV surface proteins can be profiled simultaneously with high sensitivity and specificity. This approach was also successfully applied to similar protocol using cell and serum samples. We further described the development of a micro-size exclusion chromatography method, where we were able to detect EV surface proteins with as little as 10 μL of human serum when combined with immuno-PCR. Overall, these results show that the immuno-PCR method results in rapid detection of multiple EV markers from small sample volumes in a single tube.Li SunDavid G. MeckesNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Li Sun David G. Meckes Multiplex protein profiling method for extracellular vesicle protein detection
description	Abstract Extracellular vesicles (EVs) are small nanometer-sized membrane sacs secreted into biological fluids by all cells. EVs encapsulate proteins, RNAs and metabolites from its origin cell and play important roles in intercellular communication events. Over the past decade, EVs have become a new emerging source for cancer diagnostics. One of the challenges in the study of EVs and there utility as diagnostic biomarkers is the amount of EVs needed for traditional protein analysis methods. Here, we present a new immuno-PCR method that takes advantage of commercially available TotalSeq antibodies containing DNA conjugated oligos to identify immobilized protein analysts using real-time qPCR. Using this method, we demonstrate that multiple EV surface proteins can be profiled simultaneously with high sensitivity and specificity. This approach was also successfully applied to similar protocol using cell and serum samples. We further described the development of a micro-size exclusion chromatography method, where we were able to detect EV surface proteins with as little as 10 μL of human serum when combined with immuno-PCR. Overall, these results show that the immuno-PCR method results in rapid detection of multiple EV markers from small sample volumes in a single tube.
format	article
author	Li Sun David G. Meckes
author_facet	Li Sun David G. Meckes
author_sort	Li Sun
title	Multiplex protein profiling method for extracellular vesicle protein detection
title_short	Multiplex protein profiling method for extracellular vesicle protein detection
title_full	Multiplex protein profiling method for extracellular vesicle protein detection
title_fullStr	Multiplex protein profiling method for extracellular vesicle protein detection
title_full_unstemmed	Multiplex protein profiling method for extracellular vesicle protein detection
title_sort	multiplex protein profiling method for extracellular vesicle protein detection
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/9134c49c675a4c8c9f16183c8492ad6a
work_keys_str_mv	AT lisun multiplexproteinprofilingmethodforextracellularvesicleproteindetection AT davidgmeckes multiplexproteinprofilingmethodforextracellularvesicleproteindetection
_version_	1718379658066526208

Multiplex protein profiling method for extracellular vesicle protein detection

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