Enrichment of extracellular vesicle subpopulations via affinity chromatography

Michelle E. Hung, Stephen B. Lenzini, Devin M. Stranford, Joshua N Leonard*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)

Abstract

Extracellular vesicles (EVs) are secreted nanoscale particles that transfer biomolecular cargo between cells in multicellular organisms. EVs play a variety of roles in intercellular communication and are being explored as potential vehicles for delivery of therapeutic biomolecules. However, EVs are highly heterogeneous in composition and biogenesis route, and this poses substantial challenges for understanding the role of EVs in biology and for harnessing these mechanisms for therapeutic applications, for which purifying therapeutic EVs from mixed EV populations may be necessary. Currently, technologies for isolating EV subsets are limited by overlapping physical properties among EV subsets. To meet this need, here we report an affinity chromatography-based method for enriching a specific EV subset from a heterogeneous EV starting population. By displaying an affinity tagged protein (tag-protein) on the EV surface, tagged EVs may be specifically isolated using simple affinity chromatography. Moreover, recovered EVs are enriched in the tag-protein relative to the starting population of EVs and relative to EVs purified from cell culture supernatant by standard differential centrifugation. Furthermore, chromatographically enriched EVs confer enhanced delivery of a cargo protein to recipient cells (via enhancing the amount of cargo protein per EV) relative to EVs isolated by centrifugation. Altogether, affinity chromatographic enrichment of EV subsets is a viable and facile strategy for investigating EV biology and for harnessing EVs for therapeutic applications.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc
Pages109-124
Number of pages16
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1740
ISSN (Print)1064-3745

Fingerprint

Affinity Chromatography
Extracellular Vesicles
Centrifugation
Proteins
Population
Therapeutics

Keywords

  • Chromatography
  • Exosome
  • Microvesicle
  • Purification
  • Vesicle

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Hung, M. E., Lenzini, S. B., Stranford, D. M., & Leonard, J. N. (2018). Enrichment of extracellular vesicle subpopulations via affinity chromatography. In Methods in Molecular Biology (pp. 109-124). (Methods in Molecular Biology; Vol. 1740). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7652-2_9
Hung, Michelle E. ; Lenzini, Stephen B. ; Stranford, Devin M. ; Leonard, Joshua N. / Enrichment of extracellular vesicle subpopulations via affinity chromatography. Methods in Molecular Biology. Humana Press Inc, 2018. pp. 109-124 (Methods in Molecular Biology).
@inbook{814cb384ac944f71a2a3aa4edf245434,
title = "Enrichment of extracellular vesicle subpopulations via affinity chromatography",
abstract = "Extracellular vesicles (EVs) are secreted nanoscale particles that transfer biomolecular cargo between cells in multicellular organisms. EVs play a variety of roles in intercellular communication and are being explored as potential vehicles for delivery of therapeutic biomolecules. However, EVs are highly heterogeneous in composition and biogenesis route, and this poses substantial challenges for understanding the role of EVs in biology and for harnessing these mechanisms for therapeutic applications, for which purifying therapeutic EVs from mixed EV populations may be necessary. Currently, technologies for isolating EV subsets are limited by overlapping physical properties among EV subsets. To meet this need, here we report an affinity chromatography-based method for enriching a specific EV subset from a heterogeneous EV starting population. By displaying an affinity tagged protein (tag-protein) on the EV surface, tagged EVs may be specifically isolated using simple affinity chromatography. Moreover, recovered EVs are enriched in the tag-protein relative to the starting population of EVs and relative to EVs purified from cell culture supernatant by standard differential centrifugation. Furthermore, chromatographically enriched EVs confer enhanced delivery of a cargo protein to recipient cells (via enhancing the amount of cargo protein per EV) relative to EVs isolated by centrifugation. Altogether, affinity chromatographic enrichment of EV subsets is a viable and facile strategy for investigating EV biology and for harnessing EVs for therapeutic applications.",
keywords = "Chromatography, Exosome, Microvesicle, Purification, Vesicle",
author = "Hung, {Michelle E.} and Lenzini, {Stephen B.} and Stranford, {Devin M.} and Leonard, {Joshua N}",
year = "2018",
month = "1",
day = "1",
doi = "10.1007/978-1-4939-7652-2_9",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc",
pages = "109--124",
booktitle = "Methods in Molecular Biology",

}

Hung, ME, Lenzini, SB, Stranford, DM & Leonard, JN 2018, Enrichment of extracellular vesicle subpopulations via affinity chromatography. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1740, Humana Press Inc, pp. 109-124. https://doi.org/10.1007/978-1-4939-7652-2_9

Enrichment of extracellular vesicle subpopulations via affinity chromatography. / Hung, Michelle E.; Lenzini, Stephen B.; Stranford, Devin M.; Leonard, Joshua N.

Methods in Molecular Biology. Humana Press Inc, 2018. p. 109-124 (Methods in Molecular Biology; Vol. 1740).

Research output: Chapter in Book/Report/Conference proceedingChapter

TY - CHAP

T1 - Enrichment of extracellular vesicle subpopulations via affinity chromatography

AU - Hung, Michelle E.

AU - Lenzini, Stephen B.

AU - Stranford, Devin M.

AU - Leonard, Joshua N

PY - 2018/1/1

Y1 - 2018/1/1

N2 - Extracellular vesicles (EVs) are secreted nanoscale particles that transfer biomolecular cargo between cells in multicellular organisms. EVs play a variety of roles in intercellular communication and are being explored as potential vehicles for delivery of therapeutic biomolecules. However, EVs are highly heterogeneous in composition and biogenesis route, and this poses substantial challenges for understanding the role of EVs in biology and for harnessing these mechanisms for therapeutic applications, for which purifying therapeutic EVs from mixed EV populations may be necessary. Currently, technologies for isolating EV subsets are limited by overlapping physical properties among EV subsets. To meet this need, here we report an affinity chromatography-based method for enriching a specific EV subset from a heterogeneous EV starting population. By displaying an affinity tagged protein (tag-protein) on the EV surface, tagged EVs may be specifically isolated using simple affinity chromatography. Moreover, recovered EVs are enriched in the tag-protein relative to the starting population of EVs and relative to EVs purified from cell culture supernatant by standard differential centrifugation. Furthermore, chromatographically enriched EVs confer enhanced delivery of a cargo protein to recipient cells (via enhancing the amount of cargo protein per EV) relative to EVs isolated by centrifugation. Altogether, affinity chromatographic enrichment of EV subsets is a viable and facile strategy for investigating EV biology and for harnessing EVs for therapeutic applications.

AB - Extracellular vesicles (EVs) are secreted nanoscale particles that transfer biomolecular cargo between cells in multicellular organisms. EVs play a variety of roles in intercellular communication and are being explored as potential vehicles for delivery of therapeutic biomolecules. However, EVs are highly heterogeneous in composition and biogenesis route, and this poses substantial challenges for understanding the role of EVs in biology and for harnessing these mechanisms for therapeutic applications, for which purifying therapeutic EVs from mixed EV populations may be necessary. Currently, technologies for isolating EV subsets are limited by overlapping physical properties among EV subsets. To meet this need, here we report an affinity chromatography-based method for enriching a specific EV subset from a heterogeneous EV starting population. By displaying an affinity tagged protein (tag-protein) on the EV surface, tagged EVs may be specifically isolated using simple affinity chromatography. Moreover, recovered EVs are enriched in the tag-protein relative to the starting population of EVs and relative to EVs purified from cell culture supernatant by standard differential centrifugation. Furthermore, chromatographically enriched EVs confer enhanced delivery of a cargo protein to recipient cells (via enhancing the amount of cargo protein per EV) relative to EVs isolated by centrifugation. Altogether, affinity chromatographic enrichment of EV subsets is a viable and facile strategy for investigating EV biology and for harnessing EVs for therapeutic applications.

KW - Chromatography

KW - Exosome

KW - Microvesicle

KW - Purification

KW - Vesicle

UR - http://www.scopus.com/inward/record.url?scp=85041733961&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85041733961&partnerID=8YFLogxK

U2 - 10.1007/978-1-4939-7652-2_9

DO - 10.1007/978-1-4939-7652-2_9

M3 - Chapter

T3 - Methods in Molecular Biology

SP - 109

EP - 124

BT - Methods in Molecular Biology

PB - Humana Press Inc

ER -

Hung ME, Lenzini SB, Stranford DM, Leonard JN. Enrichment of extracellular vesicle subpopulations via affinity chromatography. In Methods in Molecular Biology. Humana Press Inc. 2018. p. 109-124. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-7652-2_9