Infrared and Raman spectroscopy for purity assessment of extracellular vesicles

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Zini , J , Saari , H , Ciana , P , Viitala , T , Lohmus , A , Saarinen , J & Yliperttula , M 2022 , ' Infrared and Raman spectroscopy for purity assessment of extracellular vesicles ' , European Journal of Pharmaceutical Sciences , vol. 172 , 106135 .

Title: Infrared and Raman spectroscopy for purity assessment of extracellular vesicles
Author: Zini, Jacopo; Saari, Heikki; Ciana, Paolo; Viitala, Tapani; Lohmus, Andres; Saarinen, Jukka; Yliperttula, Marjo
Contributor organization: Division of Pharmaceutical Biosciences
Biopharmaceutics Group
Molecular and Integrative Biosciences Research Programme
Divisions of Faculty of Pharmacy
Division of Pharmaceutical Chemistry and Technology
Drug Research Program
Pharmaceutical biophysics group
Date: 2022-05-01
Language: eng
Number of pages: 12
Belongs to series: European Journal of Pharmaceutical Sciences
ISSN: 0928-0987
Abstract: Extracellular vesicles (EVs) are a complex and heterogeneous population of nanoparticles involved in cell-to-cell communication. Recently, numerous studies have indicated the potential of EVs as therapeutic agents, drug carriers and diagnostic tools. However, the results of these studies are often difficult to evaluate, since different characterization methods are used to assess the purity, physical and biochemical characteristics of the EV samples. In this study, we compared four methods for the EV sample characterization and purity assessment: i) the particle-to-protein ratio based on particle analyses with nanoparticle tracking and protein concentration by bicinchoninic acid assay, ii) Western Blot analysis for specific EV biomarkers, iii) two spectroscopic lipid-to protein ratios by either the attenuated total reflection Fourier transform infrared (ATR-FTIR) or Raman spectroscopy. The results confirm the value of Raman and ATR-FTIR spectroscopy as robust, fast and operator independent tools that require only a few microliters of EV sample. We propose that the spectroscopic lipid-to protein (Li/Pr) ratios are reliable parameters for the purity assessment of EV preparations. Moreover, apart from determining protein concentrations, we show that ATR-FTIR spectroscopy can also be used for indirect measurements of EV concentrations. Nevertheless, the Li/Pr ratios do not represent full characterization of the EV preparations. For a complete characterization of selected EV preparations, we recommend also additional use of particle size distribution and EV biomarker analysis.
Subject: Extracellular vesicles
Purification protocols
Raman spectroscopy
IR spectroscopy
Size exclusion chromatography
Differential centrifugation
Density gradient
Nanoparticle tracking analysis
1182 Biochemistry, cell and molecular biology
318 Medical biotechnology
Peer reviewed: Yes
Rights: cc_by
Usage restriction: openAccess
Self-archived version: publishedVersion

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