Skip to main content
SpringerLink
Log in

A Microfluidic SERS Assay to Characterize the Phenotypic Heterogeneity in Cancer-Derived Small Extracellular Vesicles

  • Protocol
  • First Online:
Microfluidic Systems for Cancer Diagnosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2679))

  • 562 Accesses

Abstract

Small extracellular vesicles (sEVs) are nanoscopic bioparticles that transport biomolecular cargoes between cells. sEVs have been implicated in many pathological processes such as cancer, rendering them as promising targets for therapeutics and diagnostics. Characterizing phenotypic differences in sEV biomolecular cargos could support understanding their roles in cancer. However, this is difficult due to similar physical properties of sEVs and requirement for highly sensitive analysis. Our method describes the preparation and operation of a microfluidic immunoassay with surface-enhanced Raman scattering (SERS) readouts, termed sEV subpopulation characterization platform (ESCP). ESCP applies an alternating current induced electrohydrodynamic flow to enhance collisions of sEVs with the antibody-functionalized sensor surface. Captured sEVs are labeled with plasmonic nanoparticles to facilitate multiplexed and highly sensitive phenotypic characterization of sEVs by SERS. ESCP is demonstrated for characterizing the expression of three tetraspanins (CD9, CD63, CD81) and four cancer-associated biomarkers (MCSP, MCAM, ErbB3, LNGFR) in sEVs derived from cancer cell lines and plasma samples.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 189.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 249.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Fais S, O’Driscoll L, Borras FE, Buzas E, Camussi G, Cappello F, Carvalho J, Cordeiro da Silva A, Del Portillo H, El Andaloussi S, Ficko Trček T, Furlan R, Hendrix A, Gursel I, Kralj-Iglic V, Kaeffer B, Kosanovic M, Lekka ME, Lipps G, Logozzi M, Marcilla A, Sammar M, Llorente A, Nazarenko I, Oliveira C, Pocsfalvi G, Rajendran L, Raposo G, Rohde E, Siljander P, van Niel G, Vasconcelos MH, Yáñez-Mó M, Yliperttula ML, Zarovni N, Zavec AB, Giebel B (2016) Evidence-based clinical use of nanoscale extracellular vesicles in Nanomedicine. ACS Nano 10:3886–3899

    Article  CAS  PubMed  Google Scholar 

  2. Kowal J, Arras G, Colombo M, Jouve M, Morath JP, Primdal-Bengtson B, Dingli F, Loew D, Tkach M, Théry C (2016) Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes. Proc Natl Acad Sci U S A 113:E968–E977

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Roy S, Lin H-Y, Chou C-Y, Huang C-H, Small J, Sadik N, Ayinon CM, Lansbury E, Cruz L, Yekula A, Jones PS, Balaj L, Carter BS (2019) Navigating the landscape of tumor extracellular vesicle heterogeneity. Int J Mol Sci 20:1349

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Lobb RJ, Becker M, Wen Wen S, Wong CS, Wiegmans AP, Leimgruber A, Möller A (2015) Optimized exosome isolation protocol for cell culture supernatant and human plasma. J extracell vesicles 4:27031

    Article  PubMed  Google Scholar 

  5. Giebel B (2017) On the function and heterogeneity of extracellular vesicles. Ann Transl Med 5:150

    Article  PubMed  PubMed Central  Google Scholar 

  6. Wang J, Wuethrich A, Lobb RJ, Antaw F, Sina AAI, Lane RE, Zhou Q, Zieschank C, Bell C, Bonazzi VF, Aoude LG, Everitt S, Yeo B, Barbour AP, Möller A, Trau M (2021) Characterizing the heterogeneity of Small extracellular vesicle populations in multiple cancer types via an ultrasensitive Chip. ACS Sens 6:3182–3194

    Article  CAS  PubMed  Google Scholar 

  7. Khondakar KR, Dey S, Wuethrich A, Sina AAI, Trau M (2019) Toward personalized cancer treatment: from diagnostics to therapy monitoring in miniaturized electrohydrodynamic systems. Acc Chem Res 52:2113–2123

    Article  CAS  PubMed  Google Scholar 

  8. Zhao J, Lui H, McLean DI, Zeng H (2007) Automated autofluorescence background subtraction algorithm for biomedical Raman spectroscopy. Appl Spectrosc 61:1225–1232

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

M.T. and A.W. acknowledge the support by the Australian Research Council (DP210103151) and the National Breast Cancer Foundation of Australia (CG-12-07), National Health and Medical Research Council (APP1175047 and APP1185907), and Cancer Australia (AppID_2010799). J.W. is grateful for the support by the National Natural Science Foundation of China (No. 12074069).

Author information

Authors and Affiliations

  1. Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Provincial Key Laboratory of Photonics Technology, Fujian Normal University, Fuzhou, China

    Jing Wang

  2. Centre for Personalized Nanomedicine, Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, QLD, Australia

    Matt Trau & Alain Wuethrich

  3. School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD, Australia

    Matt Trau

Authors
  1. Jing Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Matt Trau
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alain Wuethrich
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Alain Wuethrich .

Editor information

Editors and Affiliations

  1. Institute of Human Biology (IHB), Roche Pharma Research and Early Development, Roche Innovation Center, Basel, Switzerland

    Jose L. Garcia-Cordero

  2. Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, MN, USA

    Alexander Revzin

Rights and permissions

Reprints and permissions

Copyright information

© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Wang, J., Trau, M., Wuethrich, A. (2023). A Microfluidic SERS Assay to Characterize the Phenotypic Heterogeneity in Cancer-Derived Small Extracellular Vesicles. In: Garcia-Cordero, J.L., Revzin, A. (eds) Microfluidic Systems for Cancer Diagnosis . Methods in Molecular Biology, vol 2679. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3271-0_17

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-3271-0_17

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3270-3

  • Online ISBN: 978-1-0716-3271-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation