Surface Enhanced Raman Scattering of Extracellular Vesicles for Cancer Diagnostics Despite Isolation Dependent Lipoprotein Contamination

Overview

Journal Nanoscale

Specialty Biotechnology

Date 2021 Sep 2

PMID 34473170

Citations 21

Authors

Hanna J Koster

Tatu Rojalin

Alyssa Powell

Dina Pham

Rachel R Mizenko

Andrew C Birkeland

Randy P Carney

Affiliations

Soon will be listed here.

Abstract

Given the emerging diagnostic utility of extracellular vesicles (EVs), it is important to account for non-EV contaminants. Lipoprotein present in EV-enriched isolates may inflate particle counts and decrease sensitivity to biomarkers of interest, skewing chemical analyses and perpetuating downstream issues in labeling or functional analysis. Using label free surface enhanced Raman scattering (SERS), we confirm that three common EV isolation methods (differential ultracentrifugation, density gradient ultracentrifugation, and size exclusion chromatography) yield variable lipoprotein content. We demonstrate that a dual-isolation method is necessary to isolate EVs from the major classes of lipoprotein. However, combining SERS analysis with machine learning assisted classification, we show that the disease state is the main driver of distinction between EV samples, and largely unaffected by choice of isolation. Ultimately, this study describes a convenient SERS assay to retain accurate diagnostic information from clinical samples by overcoming differences in lipoprotein contamination according to isolation method.

Citing Articles

Shining a light on fluorescent EV dyes: Evaluating efficacy, specificity and suitability by nano-flow cytometry.

Brealey J, Lees R, Tempest R, Law A, Guarnerio S, Maani R J Extracell Biol. 2024; 3(10):e70006.

PMID: 39399294 PMC: 11465455. DOI: 10.1002/jex2.70006.

A critical systematic review of extracellular vesicle clinical trials.

Mizenko R, Feaver M, Bozkurt B, Lowe N, Nguyen B, Huang K J Extracell Vesicles. 2024; 13(10):e12510.

PMID: 39330928 PMC: 11428870. DOI: 10.1002/jev2.12510.

Plasma-derived extracellular vesicles (EVs) as biomarkers of sepsis in burn patients via label-free Raman spectroscopy.

OToole H, Lowe N, Arun V, Kolesov A, Palmieri T, Tran N J Extracell Vesicles. 2024; 13(9):e12506.

PMID: 39300768 PMC: 11529045. DOI: 10.1002/jev2.12506.

Extracellular vesicles may provide an alternative detoxification pathway during skeletal muscle myoblast ageing.

Fernandez-Rhodes M, Buchan E, Gagnon S, Qian J, Gethings L, Lees R J Extracell Biol. 2024; 3(8):e171.

PMID: 39169919 PMC: 11336379. DOI: 10.1002/jex2.171.

Deep autoencoder as an interpretable tool for Raman spectroscopy investigation of chemical and extracellular vesicle mixtures.

Kazemzadeh M, Martinez-Calderon M, Otupiri R, Artuyants A, Lowe M, Ning X Biomed Opt Express. 2024; 15(7):4220-4236.

PMID: 39022543 PMC: 11249694. DOI: 10.1364/BOE.522376.

References

Boucher J, Nguyen T, Sparks D . Lipoprotein electrostatic properties regulate hepatic lipase association and activity. Biochem Cell Biol. 2007; 85(6):696-708. DOI: 10.1139/o07-137. View

Driedonks T, Nolte-t Hoen E . Circulating Y-RNAs in Extracellular Vesicles and Ribonucleoprotein Complexes; Implications for the Immune System. Front Immunol. 2019; 9:3164. PMC: 6340977. DOI: 10.3389/fimmu.2018.03164. View

Onodi Z, Pelyhe C, Nagy C, Brenner G, Almasi L, Kittel A . Isolation of High-Purity Extracellular Vesicles by the Combination of Iodixanol Density Gradient Ultracentrifugation and Bind-Elute Chromatography From Blood Plasma. Front Physiol. 2018; 9:1479. PMC: 6206048. DOI: 10.3389/fphys.2018.01479. View

Shin H, Seo D, Choi Y . Extracellular Vesicle Identification Using Label-Free Surface-Enhanced Raman Spectroscopy: Detection and Signal Analysis Strategies. Molecules. 2020; 25(21). PMC: 7664897. DOI: 10.3390/molecules25215209. View

Cvjetkovic A, Lotvall J, Lasser C . The influence of rotor type and centrifugation time on the yield and purity of extracellular vesicles. J Extracell Vesicles. 2014; 3. PMC: 3967015. DOI: 10.3402/jev.v3.23111. View

Baranyai T, Herczeg K, Onodi Z, Voszka I, Modos K, Marton N . Isolation of Exosomes from Blood Plasma: Qualitative and Quantitative Comparison of Ultracentrifugation and Size Exclusion Chromatography Methods. PLoS One. 2015; 10(12):e0145686. PMC: 4686892. DOI: 10.1371/journal.pone.0145686. View

Thery C, Witwer K, Aikawa E, Alcaraz M, Anderson J, Andriantsitohaina R . Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles. 2019; 7(1):1535750. PMC: 6322352. DOI: 10.1080/20013078.2018.1535750. View

Zonneveld M, Brisson A, van Herwijnen M, Tan S, van de Lest C, Redegeld F . Recovery of extracellular vesicles from human breast milk is influenced by sample collection and vesicle isolation procedures. J Extracell Vesicles. 2014; 3. PMC: 4139932. DOI: 10.3402/jev.v3.24215. View

Fruhbeis C, Helmig S, Tug S, Simon P, Kramer-Albers E . Physical exercise induces rapid release of small extracellular vesicles into the circulation. J Extracell Vesicles. 2015; 4:28239. PMC: 4491306. DOI: 10.3402/jev.v4.28239. View

10.

Jeng M, Sharma M, Sharma L, Chao T, Huang S, Chang L . Raman Spectroscopy Analysis for Optical Diagnosis of Oral Cancer Detection. J Clin Med. 2019; 8(9). PMC: 6780219. DOI: 10.3390/jcm8091313. View

11.

Yuana Y, Levels J, Grootemaat A, Sturk A, Nieuwland R . Co-isolation of extracellular vesicles and high-density lipoproteins using density gradient ultracentrifugation. J Extracell Vesicles. 2014; 3. PMC: 4090368. DOI: 10.3402/jev.v3.23262. View

12.

Kelly J, Trevisan J, Scott A, Carmichael P, Pollock H, Martin-Hirsch P . Biospectroscopy to metabolically profile biomolecular structure: a multistage approach linking computational analysis with biomarkers. J Proteome Res. 2011; 10(4):1437-48. DOI: 10.1021/pr101067u. View

13.

Stremersch S, Marro M, Pinchasik B, Baatsen P, Hendrix A, De Smedt S . Identification of Individual Exosome-Like Vesicles by Surface Enhanced Raman Spectroscopy. Small. 2016; 12(24):3292-301. DOI: 10.1002/smll.201600393. View

14.

Ma Y, Jiang L, Mei Y, Song R, Tian D, Huang H . Colorimetric sensing strategy for mercury(II) and melamine utilizing cysteamine-modified gold nanoparticles. Analyst. 2013; 138(18):5338-43. DOI: 10.1039/c3an00690e. View

15.

Feng S, Lin D, Lin J, Li B, Huang Z, Chen G . Blood plasma surface-enhanced Raman spectroscopy for non-invasive optical detection of cervical cancer. Analyst. 2013; 138(14):3967-74. DOI: 10.1039/c3an36890d. View

16.

Arraud N, Linares R, Tan S, Gounou C, Pasquet J, Mornet S . Extracellular vesicles from blood plasma: determination of their morphology, size, phenotype and concentration. J Thromb Haemost. 2014; 12(5):614-27. DOI: 10.1111/jth.12554. View

17.

Carney R, Hazari S, Rojalin T, Knudson A, Gao T, Tang Y . Targeting Tumor-Associated Exosomes with Integrin-Binding Peptides. Adv Biosyst. 2018; 1(5). PMC: 6001286. DOI: 10.1002/adbi.201600038. View

18.

Mol E, Goumans M, Doevendans P, Sluijter J, Vader P . Higher functionality of extracellular vesicles isolated using size-exclusion chromatography compared to ultracentrifugation. Nanomedicine. 2017; 13(6):2061-2065. DOI: 10.1016/j.nano.2017.03.011. View

19.

Boing A, van der Pol E, Grootemaat A, Coumans F, Sturk A, Nieuwland R . Single-step isolation of extracellular vesicles by size-exclusion chromatography. J Extracell Vesicles. 2014; 3. PMC: 4159761. DOI: 10.3402/jev.v3.23430. View

20.

Klein-Scory S, Tehrani M, Eilert-Micus C, Adamczyk K, Wojtalewicz N, Schnolzer M . New insights in the composition of extracellular vesicles from pancreatic cancer cells: implications for biomarkers and functions. Proteome Sci. 2014; 12(1):50. PMC: 4251850. DOI: 10.1186/s12953-014-0050-5. View