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Affinity-based isolation of extracellular vesicles by means of single-domain antibodies bound to macroporous methacrylate-based copolymer

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2022
Affinity-based_isolation.pdf (5.499Mb)
Authors
Filipović, Lidija
Spasojević, Milica
Prodanović, Radivoje
Korać, Aleksandra
Matijaševic, Suzana
Brajušković, Goran
de Marco, Ario
Popović, Milica
Article (Published version)
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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 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 / Nanobodies
Source:
New Biotechnology, 2022, 69, 36-48
Publisher:
  • Elsevier
Funding / projects:
  • Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 200168 (University of Belgrade, Faculty of Chemistry) (RS-200168)
  • Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 200288 (Innovation Center of the Faculty of Chemistry) (RS-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

Scopus: 2-s2.0-85126528334
[ Google Scholar ]
URI
https://pubmed.ncbi.nlm.nih.gov/35301156/
http://cherry.chem.bg.ac.rs/handle/123456789/5152
Collections
  • Publikacije
  • Publikacije
Institution/Community
Hemijski fakultet
TY  - 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
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",
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.. (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 M. 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, "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 . .

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