FO‐SPR biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matrices
Abstract Extracellular vesicles (EVs) have drawn huge attention for diagnosing myriad of diseases, including cancer. However, the EV detection and analyses procedures often lack much desired sample standardization. To address this, we used well‐characterized recombinant EVs (rEVs) for the first time...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2021-02-01
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| Series: | Journal of Extracellular Vesicles |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/jev2.12059 |
| _version_ | 1819125169028857856 |
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| author | Yagmur Yildizhan Venkata Suresh Vajrala Edward Geeurickx Charles Declerck Nevena Duskunovic Delphine De Sutter Sam Noppen Filip Delport Dominique Schols Johannes V. Swinnen Sven Eyckerman An Hendrix Jeroen Lammertyn Dragana Spasic |
| author_facet | Yagmur Yildizhan Venkata Suresh Vajrala Edward Geeurickx Charles Declerck Nevena Duskunovic Delphine De Sutter Sam Noppen Filip Delport Dominique Schols Johannes V. Swinnen Sven Eyckerman An Hendrix Jeroen Lammertyn Dragana Spasic |
| author_sort | Yagmur Yildizhan |
| collection | DOAJ |
| description | Abstract Extracellular vesicles (EVs) have drawn huge attention for diagnosing myriad of diseases, including cancer. However, the EV detection and analyses procedures often lack much desired sample standardization. To address this, we used well‐characterized recombinant EVs (rEVs) for the first time as a biological reference material in developing a fiber optic surface plasmon resonance (FO‐SPR) bioassay. In this context, EV binding on the FO‐SPR probes was achieved only with EV‐specific antibodies (e.g. anti‐CD9 and anti‐CD63) but not with non‐specific anti‐IgG. To increase detection sensitivity, we tested six different combinations of EV‐specific antibodies in a sandwich bioassay. Calibration curves were generated with two most effective combinations (anti‐CD9/Banti‐CD81 and anti‐CD63/Banti‐CD9), resulting in 103 and 104 times higher sensitivity than the EV concentration in human blood plasma from healthy or cancer patients, respectively. Additionally, by using anti‐CD63/Banti‐CD9, we detected rEVs spiked in cell culture medium and HEK293 endogenous EVs in the same matrix without any prior EV purification or enrichment. Lastly, we selectively captured breast cancer cell EVs spiked in blood plasma using anti‐EpCAM antibody on the FO‐SPR surface. The obtained results combined with FO‐SPR real‐time monitoring, fast response time and ease of operation, demonstrate its outstanding potential for EV quantification and analysis. |
| first_indexed | 2024-12-22T07:35:52Z |
| format | Article |
| id | doaj.art-4e8411d58d5f41989eb27ca1ef801402 |
| institution | Directory Open Access Journal |
| issn | 2001-3078 |
| language | English |
| last_indexed | 2024-12-22T07:35:52Z |
| publishDate | 2021-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Extracellular Vesicles |
| spelling | doaj.art-4e8411d58d5f41989eb27ca1ef8014022022-12-21T18:33:53ZengWileyJournal of Extracellular Vesicles2001-30782021-02-01104n/an/a10.1002/jev2.12059FO‐SPR biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matricesYagmur Yildizhan0Venkata Suresh Vajrala1Edward Geeurickx2Charles Declerck3Nevena Duskunovic4Delphine De Sutter5Sam Noppen6Filip Delport7Dominique Schols8Johannes V. Swinnen9Sven Eyckerman10An Hendrix11Jeroen Lammertyn12Dragana Spasic13Department of Biosystems Biosensors group, KU Leuven Leuven BelgiumDepartment of Biosystems Biosensors group, KU Leuven Leuven BelgiumDepartment of Human Structure and Repair Laboratory of Experimental Cancer Research Ghent University Ghent BelgiumDepartment of Biosystems Biosensors group, KU Leuven Leuven BelgiumDepartment of Biosystems Biosensors group, KU Leuven Leuven BelgiumVIB Center for Medical Biotechnology & Department of Biomolecular Medicine Ghent University GhentDepartment of Microbiology Immunology and Transplantation, Laboratory of Virology and Chemotherapy, Rega Institute KU Leuven Leuven BelgiumFOx Biosystems Bioville Diepenbeek BelgiumDepartment of Microbiology Immunology and Transplantation, Laboratory of Virology and Chemotherapy, Rega Institute KU Leuven Leuven BelgiumDepartment of Oncology Laboratory of Lipid Metabolism and Cancer KU Leuven Leuven BelgiumVIB Center for Medical Biotechnology & Department of Biomolecular Medicine Ghent University GhentDepartment of Human Structure and Repair Laboratory of Experimental Cancer Research Ghent University Ghent BelgiumDepartment of Biosystems Biosensors group, KU Leuven Leuven BelgiumDepartment of Biosystems Biosensors group, KU Leuven Leuven BelgiumAbstract Extracellular vesicles (EVs) have drawn huge attention for diagnosing myriad of diseases, including cancer. However, the EV detection and analyses procedures often lack much desired sample standardization. To address this, we used well‐characterized recombinant EVs (rEVs) for the first time as a biological reference material in developing a fiber optic surface plasmon resonance (FO‐SPR) bioassay. In this context, EV binding on the FO‐SPR probes was achieved only with EV‐specific antibodies (e.g. anti‐CD9 and anti‐CD63) but not with non‐specific anti‐IgG. To increase detection sensitivity, we tested six different combinations of EV‐specific antibodies in a sandwich bioassay. Calibration curves were generated with two most effective combinations (anti‐CD9/Banti‐CD81 and anti‐CD63/Banti‐CD9), resulting in 103 and 104 times higher sensitivity than the EV concentration in human blood plasma from healthy or cancer patients, respectively. Additionally, by using anti‐CD63/Banti‐CD9, we detected rEVs spiked in cell culture medium and HEK293 endogenous EVs in the same matrix without any prior EV purification or enrichment. Lastly, we selectively captured breast cancer cell EVs spiked in blood plasma using anti‐EpCAM antibody on the FO‐SPR surface. The obtained results combined with FO‐SPR real‐time monitoring, fast response time and ease of operation, demonstrate its outstanding potential for EV quantification and analysis.https://doi.org/10.1002/jev2.12059biosensorcomplex matricesextracellular vesiclesfiber opticssurface plasmon resonance |
| spellingShingle | Yagmur Yildizhan Venkata Suresh Vajrala Edward Geeurickx Charles Declerck Nevena Duskunovic Delphine De Sutter Sam Noppen Filip Delport Dominique Schols Johannes V. Swinnen Sven Eyckerman An Hendrix Jeroen Lammertyn Dragana Spasic FO‐SPR biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matrices Journal of Extracellular Vesicles biosensor complex matrices extracellular vesicles fiber optics surface plasmon resonance |
| title | FO‐SPR biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matrices |
| title_full | FO‐SPR biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matrices |
| title_fullStr | FO‐SPR biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matrices |
| title_full_unstemmed | FO‐SPR biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matrices |
| title_short | FO‐SPR biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matrices |
| title_sort | fo spr biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matrices |
| topic | biosensor complex matrices extracellular vesicles fiber optics surface plasmon resonance |
| url | https://doi.org/10.1002/jev2.12059 |
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