Cargando…

Integrated immunoisolation and protein analysis of circulating exosomes using microfluidic technology

Developing blood-based tests is appealing for non-invasive disease diagnosis, especially when biopsy is difficult, costly, and sometimes not even an option. Tumor-derived exosomes have attracted increasing interest in non-invasive cancer diagnosis and monitoring of treatment response. However, the b...

Descripción completa

Detalles Bibliográficos
Autores principales: He, Mei, Crow, Jennifer, Roth, Marc, Zeng, Yong, Godwin, Andrew K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Royal Society of Chemistry 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4161194/
https://www.ncbi.nlm.nih.gov/pubmed/25099143
http://dx.doi.org/10.1039/c4lc00662c
_version_ 1782334504484995072
author He, Mei
Crow, Jennifer
Roth, Marc
Zeng, Yong
Godwin, Andrew K.
author_facet He, Mei
Crow, Jennifer
Roth, Marc
Zeng, Yong
Godwin, Andrew K.
author_sort He, Mei
collection PubMed
description Developing blood-based tests is appealing for non-invasive disease diagnosis, especially when biopsy is difficult, costly, and sometimes not even an option. Tumor-derived exosomes have attracted increasing interest in non-invasive cancer diagnosis and monitoring of treatment response. However, the biology and clinical value of exosomes remains largely unknown due in part to current technical challenges in rapid isolation, molecular classification and comprehensive analysis of exosomes. Here we developed a new microfluidic approach to streamline and expedite the exosome analysis pipeline by integrating specific immunoisolation and targeted protein analysis of circulating exosomes. Compared to the conventional methods, our approach enables selective subpopulation isolation and quantitative detection of surface and intravesicular biomarkers directly from a minimally invasive amount of plasma samples (30 μL) within ~100 min with markedly improved detection sensitivity. Using this device, we demonstrated phenotyping of exosome subpopulations by targeting a panel of common exosomal and tumor-specific markers and multiparameter analyses of intravesicular biomarkers in the selected subpopulation. We were able to assess the total expression and phosphorylation levels of IGF-1R in non-small-cell lung cancer patients by probing plasma exosomes as a non-invasive alternative to conventional tissue biopsy. We foresee that the microfluidic exosome analysis platform will form the basis for critically needed infrastructures for advancing the biology and clinical utilization of exosomes.
format Online
Article
Text
id pubmed-4161194
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher Royal Society of Chemistry
record_format MEDLINE/PubMed
spelling pubmed-41611942014-10-16 Integrated immunoisolation and protein analysis of circulating exosomes using microfluidic technology He, Mei Crow, Jennifer Roth, Marc Zeng, Yong Godwin, Andrew K. Lab Chip Chemistry Developing blood-based tests is appealing for non-invasive disease diagnosis, especially when biopsy is difficult, costly, and sometimes not even an option. Tumor-derived exosomes have attracted increasing interest in non-invasive cancer diagnosis and monitoring of treatment response. However, the biology and clinical value of exosomes remains largely unknown due in part to current technical challenges in rapid isolation, molecular classification and comprehensive analysis of exosomes. Here we developed a new microfluidic approach to streamline and expedite the exosome analysis pipeline by integrating specific immunoisolation and targeted protein analysis of circulating exosomes. Compared to the conventional methods, our approach enables selective subpopulation isolation and quantitative detection of surface and intravesicular biomarkers directly from a minimally invasive amount of plasma samples (30 μL) within ~100 min with markedly improved detection sensitivity. Using this device, we demonstrated phenotyping of exosome subpopulations by targeting a panel of common exosomal and tumor-specific markers and multiparameter analyses of intravesicular biomarkers in the selected subpopulation. We were able to assess the total expression and phosphorylation levels of IGF-1R in non-small-cell lung cancer patients by probing plasma exosomes as a non-invasive alternative to conventional tissue biopsy. We foresee that the microfluidic exosome analysis platform will form the basis for critically needed infrastructures for advancing the biology and clinical utilization of exosomes. Royal Society of Chemistry 2014-10-07 2014-08-06 /pmc/articles/PMC4161194/ /pubmed/25099143 http://dx.doi.org/10.1039/c4lc00662c Text en This journal is © The Royal Society of Chemistry 2014 http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial 3.0 Unported License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Chemistry
He, Mei
Crow, Jennifer
Roth, Marc
Zeng, Yong
Godwin, Andrew K.
Integrated immunoisolation and protein analysis of circulating exosomes using microfluidic technology
title Integrated immunoisolation and protein analysis of circulating exosomes using microfluidic technology
title_full Integrated immunoisolation and protein analysis of circulating exosomes using microfluidic technology
title_fullStr Integrated immunoisolation and protein analysis of circulating exosomes using microfluidic technology
title_full_unstemmed Integrated immunoisolation and protein analysis of circulating exosomes using microfluidic technology
title_short Integrated immunoisolation and protein analysis of circulating exosomes using microfluidic technology
title_sort integrated immunoisolation and protein analysis of circulating exosomes using microfluidic technology
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4161194/
https://www.ncbi.nlm.nih.gov/pubmed/25099143
http://dx.doi.org/10.1039/c4lc00662c
work_keys_str_mv AT hemei integratedimmunoisolationandproteinanalysisofcirculatingexosomesusingmicrofluidictechnology
AT crowjennifer integratedimmunoisolationandproteinanalysisofcirculatingexosomesusingmicrofluidictechnology
AT rothmarc integratedimmunoisolationandproteinanalysisofcirculatingexosomesusingmicrofluidictechnology
AT zengyong integratedimmunoisolationandproteinanalysisofcirculatingexosomesusingmicrofluidictechnology
AT godwinandrewk integratedimmunoisolationandproteinanalysisofcirculatingexosomesusingmicrofluidictechnology