Evaluation of small extracellular vesicles isolation methods from human serum for downstream miRNA profiling
Exosomes are a type of extracellular vesicles that carry distinct profiles of biomolecules such as lipids, proteins, DNAs, and RNAs. Despite many years of research, there is still a lack of standardized methods to isolate exosomes from clinical samples for their downstream applications. Thus, this s...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Penerbit Universiti Kebangsaan Malaysia
2025
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| Online Access: | http://journalarticle.ukm.my/25795/ http://journalarticle.ukm.my/25795/1/MA%2010%20.pdf |
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| author | Yong, Ling Sou Pei, Paulina Suu Tan Nur Afrina Muhamad Hendiri, Wickneswari Ratnam, Tilakavati Karupaiah, Chilian, William M Shamsul Mohd Zain, Sharifah Zamiah Syed Abdul Kadir, Yan, Pan Pung, Yuh-Fen |
| author_facet | Yong, Ling Sou Pei, Paulina Suu Tan Nur Afrina Muhamad Hendiri, Wickneswari Ratnam, Tilakavati Karupaiah, Chilian, William M Shamsul Mohd Zain, Sharifah Zamiah Syed Abdul Kadir, Yan, Pan Pung, Yuh-Fen |
| author_sort | Yong, Ling Sou |
| building | UKM Institutional Repository |
| collection | Online Access |
| description | Exosomes are a type of extracellular vesicles that carry distinct profiles of biomolecules such as lipids, proteins, DNAs, and RNAs. Despite many years of research, there is still a lack of standardized methods to isolate exosomes from clinical samples for their downstream applications. Thus, this study compared three different methods, which are differential ultracentrifugation (DUC), polyethylene glycol-based precipitation (PEG), and a combination of both (PEG+UC) to isolate exosomes from human serum. The isolated exosomes were evaluated by their size distribution, recovered particle concentration, particle-to-protein ratio, exosomal marker expression, and miRNA recovery. Our results indicated that all three methods successfully isolated exosomes, however, with varying yield and purity. In particular, PEG+UC produced exosomes of both high yield and high purity, DUC produced exosomes of both low yield and low purity, whereas PEG produced exosomes of high yield but low purity. Using miR-30d-5p and let-7i-5p as selected targets, our qPCR results indicated significant differences in terms of exosomal miRNA recovery between all three methods. Overall, the PEG+UC method appeared to be a less labor-intensive alternative that can isolate exosomes of both high yield and high purity from human serum without compromising the yield of miRNAs. |
| first_indexed | 2025-11-15T01:06:05Z |
| format | Article |
| id | oai:generic.eprints.org:25795 |
| institution | Universiti Kebangasaan Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T01:06:05Z |
| publishDate | 2025 |
| publisher | Penerbit Universiti Kebangsaan Malaysia |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | oai:generic.eprints.org:257952025-08-19T03:49:43Z http://journalarticle.ukm.my/25795/ Evaluation of small extracellular vesicles isolation methods from human serum for downstream miRNA profiling Yong, Ling Sou Pei, Paulina Suu Tan Nur Afrina Muhamad Hendiri, Wickneswari Ratnam, Tilakavati Karupaiah, Chilian, William M Shamsul Mohd Zain, Sharifah Zamiah Syed Abdul Kadir, Yan, Pan Pung, Yuh-Fen Exosomes are a type of extracellular vesicles that carry distinct profiles of biomolecules such as lipids, proteins, DNAs, and RNAs. Despite many years of research, there is still a lack of standardized methods to isolate exosomes from clinical samples for their downstream applications. Thus, this study compared three different methods, which are differential ultracentrifugation (DUC), polyethylene glycol-based precipitation (PEG), and a combination of both (PEG+UC) to isolate exosomes from human serum. The isolated exosomes were evaluated by their size distribution, recovered particle concentration, particle-to-protein ratio, exosomal marker expression, and miRNA recovery. Our results indicated that all three methods successfully isolated exosomes, however, with varying yield and purity. In particular, PEG+UC produced exosomes of both high yield and high purity, DUC produced exosomes of both low yield and low purity, whereas PEG produced exosomes of high yield but low purity. Using miR-30d-5p and let-7i-5p as selected targets, our qPCR results indicated significant differences in terms of exosomal miRNA recovery between all three methods. Overall, the PEG+UC method appeared to be a less labor-intensive alternative that can isolate exosomes of both high yield and high purity from human serum without compromising the yield of miRNAs. Penerbit Universiti Kebangsaan Malaysia 2025 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/25795/1/MA%2010%20.pdf Yong, Ling Sou and Pei, Paulina Suu Tan and Nur Afrina Muhamad Hendiri, and Wickneswari Ratnam, and Tilakavati Karupaiah, and Chilian, William M and Shamsul Mohd Zain, and Sharifah Zamiah Syed Abdul Kadir, and Yan, Pan and Pung, Yuh-Fen (2025) Evaluation of small extracellular vesicles isolation methods from human serum for downstream miRNA profiling. Malaysian Applied Biology, 54 (1). pp. 98-107. ISSN 0126-8643 https://jms.mabjournal.com/index.php/mab/issue/view/66 |
| spellingShingle | Yong, Ling Sou Pei, Paulina Suu Tan Nur Afrina Muhamad Hendiri, Wickneswari Ratnam, Tilakavati Karupaiah, Chilian, William M Shamsul Mohd Zain, Sharifah Zamiah Syed Abdul Kadir, Yan, Pan Pung, Yuh-Fen Evaluation of small extracellular vesicles isolation methods from human serum for downstream miRNA profiling |
| title | Evaluation of small extracellular vesicles isolation methods from human serum for downstream miRNA profiling |
| title_full | Evaluation of small extracellular vesicles isolation methods from human serum for downstream miRNA profiling |
| title_fullStr | Evaluation of small extracellular vesicles isolation methods from human serum for downstream miRNA profiling |
| title_full_unstemmed | Evaluation of small extracellular vesicles isolation methods from human serum for downstream miRNA profiling |
| title_short | Evaluation of small extracellular vesicles isolation methods from human serum for downstream miRNA profiling |
| title_sort | evaluation of small extracellular vesicles isolation methods from human serum for downstream mirna profiling |
| url | http://journalarticle.ukm.my/25795/ http://journalarticle.ukm.my/25795/ http://journalarticle.ukm.my/25795/1/MA%2010%20.pdf |