Affinity-based isolation of extracellular vesicles by means of single-domain antibodies bound to macroporous methacrylate-based copolymer
Authors
Filipović, LidijaSpasojević, Milica
Prodanović, Radivoje
Korać, Aleksandra
Matijaševic, Suzana
Brajušković, Goran
de Marco, Ario
Popović, Milica M.
Article (Published version)
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Show full item recordAbstract
Correct elucidation of physiological and pathological processes mediated by extracellular vesicles (EV) is highly dependent on the reliability of the method used for their purification. Currently available chemical/physical protocols for sample fractionation are time-consuming, often scarcely reproducible and their yields are low. Immuno-capture based approaches could represent an effective purification alternative to obtain homogeneous EV samples. An easy-to-operate chromatography system was set-up for the purification of intact EVs based on a single domain (VHH) antibodies-copolymer matrix suitable for biological samples as different as conditioned cell culture medium and human plasma. Methacrylate-based copolymer is a porous solid support, the chemical versatility of which enables its efficient functionalization with VHHs. The combined analyses of morphological features and biomarker (CD9, CD63 and CD81) presence indicated that the recovered EVs were exosomes. The lipoprotein marker...s APO-A1 and APO-B were both negative in tested samples. This is the first report demonstrating the successful application of spherical porous methacrylate-based copolymer coupled with VHHs for the exosome isolation from biological fluids. This inexpensive immunoaffinity method has the potential to be applied for the isolation of EVs belonging to different morphological and physiological classes.
Keywords:
Exosomes / Extracellular vesicles / Immunopurification / Methacrylate-based copolymer / NanobodiesSource:
New Biotechnology, 2022, 69, 36-48Publisher:
- Elsevier
Funding / projects:
- Ministry of Science, Technological Development and Innovation of the Republic of Serbia, institutional funding - 200168 (University of Belgrade, Faculty of Chemistry) (RS-MESTD-inst-2020-200168)
- Ministry of Science, Technological Development and Innovation of the Republic of Serbia, institutional funding - 200288 (Innovation Center of the Faculty of Chemistry) (RS-MESTD-inst-2020-200288)
Note:
- Supplementary material: https://cherry.chem.bg.ac.rs/handle/123456789/5153
Related info:
- Referenced by
https://cherry.chem.bg.ac.rs/handle/123456789/5153
DOI: 10.1016/j.nbt.2022.03.001
ISSN: 1871-6784
WoS: 00079129900000
Scopus: 2-s2.0-85126528334
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Institution/Community
Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Filipović, Lidija AU - Spasojević, Milica AU - Prodanović, Radivoje AU - Korać, Aleksandra AU - Matijaševic, Suzana AU - Brajušković, Goran AU - de Marco, Ario AU - Popović, Milica M. PY - 2022 UR - https://pubmed.ncbi.nlm.nih.gov/35301156/ UR - http://cherry.chem.bg.ac.rs/handle/123456789/5152 AB - Correct elucidation of physiological and pathological processes mediated by extracellular vesicles (EV) is highly dependent on the reliability of the method used for their purification. Currently available chemical/physical protocols for sample fractionation are time-consuming, often scarcely reproducible and their yields are low. Immuno-capture based approaches could represent an effective purification alternative to obtain homogeneous EV samples. An easy-to-operate chromatography system was set-up for the purification of intact EVs based on a single domain (VHH) antibodies-copolymer matrix suitable for biological samples as different as conditioned cell culture medium and human plasma. Methacrylate-based copolymer is a porous solid support, the chemical versatility of which enables its efficient functionalization with VHHs. The combined analyses of morphological features and biomarker (CD9, CD63 and CD81) presence indicated that the recovered EVs were exosomes. The lipoprotein markers APO-A1 and APO-B were both negative in tested samples. This is the first report demonstrating the successful application of spherical porous methacrylate-based copolymer coupled with VHHs for the exosome isolation from biological fluids. This inexpensive immunoaffinity method has the potential to be applied for the isolation of EVs belonging to different morphological and physiological classes. PB - Elsevier T2 - New Biotechnology T1 - Affinity-based isolation of extracellular vesicles by means of single-domain antibodies bound to macroporous methacrylate-based copolymer VL - 69 SP - 36 EP - 48 DO - 10.1016/j.nbt.2022.03.001 ER -
@article{ author = "Filipović, Lidija and Spasojević, Milica and Prodanović, Radivoje and Korać, Aleksandra and Matijaševic, Suzana and Brajušković, Goran and de Marco, Ario and Popović, Milica M.", year = "2022", abstract = "Correct elucidation of physiological and pathological processes mediated by extracellular vesicles (EV) is highly dependent on the reliability of the method used for their purification. Currently available chemical/physical protocols for sample fractionation are time-consuming, often scarcely reproducible and their yields are low. Immuno-capture based approaches could represent an effective purification alternative to obtain homogeneous EV samples. An easy-to-operate chromatography system was set-up for the purification of intact EVs based on a single domain (VHH) antibodies-copolymer matrix suitable for biological samples as different as conditioned cell culture medium and human plasma. Methacrylate-based copolymer is a porous solid support, the chemical versatility of which enables its efficient functionalization with VHHs. The combined analyses of morphological features and biomarker (CD9, CD63 and CD81) presence indicated that the recovered EVs were exosomes. The lipoprotein markers APO-A1 and APO-B were both negative in tested samples. This is the first report demonstrating the successful application of spherical porous methacrylate-based copolymer coupled with VHHs for the exosome isolation from biological fluids. This inexpensive immunoaffinity method has the potential to be applied for the isolation of EVs belonging to different morphological and physiological classes.", publisher = "Elsevier", journal = "New Biotechnology", title = "Affinity-based isolation of extracellular vesicles by means of single-domain antibodies bound to macroporous methacrylate-based copolymer", volume = "69", pages = "36-48", doi = "10.1016/j.nbt.2022.03.001" }
Filipović, L., Spasojević, M., Prodanović, R., Korać, A., Matijaševic, S., Brajušković, G., de Marco, A.,& Popović, M. M.. (2022). Affinity-based isolation of extracellular vesicles by means of single-domain antibodies bound to macroporous methacrylate-based copolymer. in New Biotechnology Elsevier., 69, 36-48. https://doi.org/10.1016/j.nbt.2022.03.001
Filipović L, Spasojević M, Prodanović R, Korać A, Matijaševic S, Brajušković G, de Marco A, Popović MM. Affinity-based isolation of extracellular vesicles by means of single-domain antibodies bound to macroporous methacrylate-based copolymer. in New Biotechnology. 2022;69:36-48. doi:10.1016/j.nbt.2022.03.001 .
Filipović, Lidija, Spasojević, Milica, Prodanović, Radivoje, Korać, Aleksandra, Matijaševic, Suzana, Brajušković, Goran, de Marco, Ario, Popović, Milica M., "Affinity-based isolation of extracellular vesicles by means of single-domain antibodies bound to macroporous methacrylate-based copolymer" in New Biotechnology, 69 (2022):36-48, https://doi.org/10.1016/j.nbt.2022.03.001 . .