Usefulness of Size-Exclusion Chromatography–Multi-Angle Light Scattering to Assess Particle Composition and Protein Impurities for Quality Control of Therapeutic Exosome Preparations
<b>Background:</b> Extracellular vesicles (EVs), including exosomes, are promising pharmaceutical modalities. They are purified from cell culture supernatant; however, the preparation may contain EVs with the desired therapeutic effects and different types of EVs, lipoproteins, and solub...
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2024-11-01
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| author | Hirotaka Nishimura Noritaka Hashii Tomofumi Yamamoto Yuchen Sun Takumi Miura Yoji Sato Akiko Ishii-Watabe |
| author_facet | Hirotaka Nishimura Noritaka Hashii Tomofumi Yamamoto Yuchen Sun Takumi Miura Yoji Sato Akiko Ishii-Watabe |
| author_sort | Hirotaka Nishimura |
| collection | DOAJ |
| description | <b>Background:</b> Extracellular vesicles (EVs), including exosomes, are promising pharmaceutical modalities. They are purified from cell culture supernatant; however, the preparation may contain EVs with the desired therapeutic effects and different types of EVs, lipoproteins, and soluble proteins. Evaluating the composition of particulate impurities and the levels of protein impurities in final preparations is critical for quality control. However, few analytical methods can detect these impurities. <b>Methods:</b> We established and evaluated an analytical method using size-exclusion chromatography–multi-angle light scattering (SEC-MALS) for particle and protein impurity analyses of EV samples. <b>Results:</b> In the particle size distribution analysis of EV samples, SEC-MALS showed higher resolution compared with nanoparticle tracking analysis (NTA) and dynamic light scattering (DLS). MALS showed comparable accuracy and precision to that of other methods for particle size evaluation using polystyrene standard beads with 60, 100, or 200 nm diameter. Coupling SEC-MALS with UV detection quantitatively evaluated soluble protein impurities. Proteomic analysis on the SEC-MALS-fractionated samples identified different EV and lipoprotein marker proteins in different fractions. <b>Conclusions:</b> SEC-MALS can characterize EV preparations obtained from human adipose-derived mesenchymal stem cells, suggesting that it can evaluate the particle component composition in various EV samples and therapeutic exosome preparations. |
| format | Article |
| id | doaj-art-da919ab24f9544358119c34cfd5ce93e |
| institution | OA Journals |
| issn | 1999-4923 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj-art-da919ab24f9544358119c34cfd5ce93e2025-08-20T02:01:20ZengMDPI AGPharmaceutics1999-49232024-11-011612152610.3390/pharmaceutics16121526Usefulness of Size-Exclusion Chromatography–Multi-Angle Light Scattering to Assess Particle Composition and Protein Impurities for Quality Control of Therapeutic Exosome PreparationsHirotaka Nishimura0Noritaka Hashii1Tomofumi Yamamoto2Yuchen Sun3Takumi Miura4Yoji Sato5Akiko Ishii-Watabe6Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki 210-9501, Kanagawa, JapanDivision of Biological Chemistry and Biologicals, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki 210-9501, Kanagawa, JapanDivision of Biological Chemistry and Biologicals, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki 210-9501, Kanagawa, JapanDivision of Medicinal Safety Science, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki 210-9501, Kanagawa, JapanDivision of Cell-Based Therapeutic Products, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki 210-9501, Kanagawa, JapanDivision of Drugs, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki 210-9501, Kanagawa, JapanDivision of Biological Chemistry and Biologicals, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki 210-9501, Kanagawa, Japan<b>Background:</b> Extracellular vesicles (EVs), including exosomes, are promising pharmaceutical modalities. They are purified from cell culture supernatant; however, the preparation may contain EVs with the desired therapeutic effects and different types of EVs, lipoproteins, and soluble proteins. Evaluating the composition of particulate impurities and the levels of protein impurities in final preparations is critical for quality control. However, few analytical methods can detect these impurities. <b>Methods:</b> We established and evaluated an analytical method using size-exclusion chromatography–multi-angle light scattering (SEC-MALS) for particle and protein impurity analyses of EV samples. <b>Results:</b> In the particle size distribution analysis of EV samples, SEC-MALS showed higher resolution compared with nanoparticle tracking analysis (NTA) and dynamic light scattering (DLS). MALS showed comparable accuracy and precision to that of other methods for particle size evaluation using polystyrene standard beads with 60, 100, or 200 nm diameter. Coupling SEC-MALS with UV detection quantitatively evaluated soluble protein impurities. Proteomic analysis on the SEC-MALS-fractionated samples identified different EV and lipoprotein marker proteins in different fractions. <b>Conclusions:</b> SEC-MALS can characterize EV preparations obtained from human adipose-derived mesenchymal stem cells, suggesting that it can evaluate the particle component composition in various EV samples and therapeutic exosome preparations.https://www.mdpi.com/1999-4923/16/12/1526extracellular vesiclessize exclusion chromatographymulti-angle light scatteringEV therapeuticsquality control |
| spellingShingle | Hirotaka Nishimura Noritaka Hashii Tomofumi Yamamoto Yuchen Sun Takumi Miura Yoji Sato Akiko Ishii-Watabe Usefulness of Size-Exclusion Chromatography–Multi-Angle Light Scattering to Assess Particle Composition and Protein Impurities for Quality Control of Therapeutic Exosome Preparations Pharmaceutics extracellular vesicles size exclusion chromatography multi-angle light scattering EV therapeutics quality control |
| title | Usefulness of Size-Exclusion Chromatography–Multi-Angle Light Scattering to Assess Particle Composition and Protein Impurities for Quality Control of Therapeutic Exosome Preparations |
| title_full | Usefulness of Size-Exclusion Chromatography–Multi-Angle Light Scattering to Assess Particle Composition and Protein Impurities for Quality Control of Therapeutic Exosome Preparations |
| title_fullStr | Usefulness of Size-Exclusion Chromatography–Multi-Angle Light Scattering to Assess Particle Composition and Protein Impurities for Quality Control of Therapeutic Exosome Preparations |
| title_full_unstemmed | Usefulness of Size-Exclusion Chromatography–Multi-Angle Light Scattering to Assess Particle Composition and Protein Impurities for Quality Control of Therapeutic Exosome Preparations |
| title_short | Usefulness of Size-Exclusion Chromatography–Multi-Angle Light Scattering to Assess Particle Composition and Protein Impurities for Quality Control of Therapeutic Exosome Preparations |
| title_sort | usefulness of size exclusion chromatography multi angle light scattering to assess particle composition and protein impurities for quality control of therapeutic exosome preparations |
| topic | extracellular vesicles size exclusion chromatography multi-angle light scattering EV therapeutics quality control |
| url | https://www.mdpi.com/1999-4923/16/12/1526 |
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