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FO-SPR biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matrices.
J Extracell Vesicles. 2021 02; 10(4):e12059.JE

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.

Authors+Show Affiliations

Department of Biosystems Biosensors group, KU Leuven Leuven Belgium.Department of Biosystems Biosensors group, KU Leuven Leuven Belgium.Department of Human Structure and Repair Laboratory of Experimental Cancer Research Ghent University Ghent Belgium.Department of Biosystems Biosensors group, KU Leuven Leuven Belgium.Department of Biosystems Biosensors group, KU Leuven Leuven Belgium.VIB Center for Medical Biotechnology & Department of Biomolecular Medicine Ghent University Ghent.Department of Microbiology Immunology and Transplantation, Laboratory of Virology and Chemotherapy, Rega Institute KU Leuven Leuven Belgium.FOx Biosystems Bioville Diepenbeek Belgium.Department of Microbiology Immunology and Transplantation, Laboratory of Virology and Chemotherapy, Rega Institute KU Leuven Leuven Belgium.Department of Oncology Laboratory of Lipid Metabolism and Cancer KU Leuven Leuven Belgium.VIB Center for Medical Biotechnology & Department of Biomolecular Medicine Ghent University Ghent.Department of Human Structure and Repair Laboratory of Experimental Cancer Research Ghent University Ghent Belgium.Department of Biosystems Biosensors group, KU Leuven Leuven Belgium.Department of Biosystems Biosensors group, KU Leuven Leuven Belgium.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

33664936

Citation

Yildizhan, Yagmur, et al. "FO-SPR Biosensor Calibrated With Recombinant Extracellular Vesicles Enables Specific and Sensitive Detection Directly in Complex Matrices." Journal of Extracellular Vesicles, vol. 10, no. 4, 2021, pp. e12059.
Yildizhan Y, Vajrala VS, Geeurickx E, et al. FO-SPR biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matrices. J Extracell Vesicles. 2021;10(4):e12059.
Yildizhan, Y., Vajrala, V. S., Geeurickx, E., Declerck, C., Duskunovic, N., De Sutter, D., Noppen, S., Delport, F., Schols, D., Swinnen, J. V., Eyckerman, S., Hendrix, A., Lammertyn, J., & Spasic, D. (2021). FO-SPR biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matrices. Journal of Extracellular Vesicles, 10(4), e12059. https://doi.org/10.1002/jev2.12059
Yildizhan Y, et al. FO-SPR Biosensor Calibrated With Recombinant Extracellular Vesicles Enables Specific and Sensitive Detection Directly in Complex Matrices. J Extracell Vesicles. 2021;10(4):e12059. PubMed PMID: 33664936.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - FO-SPR biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matrices. AU - Yildizhan,Yagmur, AU - Vajrala,Venkata Suresh, AU - Geeurickx,Edward, AU - Declerck,Charles, AU - Duskunovic,Nevena, AU - De Sutter,Delphine, AU - Noppen,Sam, AU - Delport,Filip, AU - Schols,Dominique, AU - Swinnen,Johannes V, AU - Eyckerman,Sven, AU - Hendrix,An, AU - Lammertyn,Jeroen, AU - Spasic,Dragana, Y1 - 2021/02/23/ PY - 2020/10/22/received PY - 2020/12/11/revised PY - 2021/01/06/accepted PY - 2021/3/5/entrez PY - 2021/3/6/pubmed PY - 2021/3/6/medline KW - biosensor KW - complex matrices KW - extracellular vesicles KW - fiber optics KW - surface plasmon resonance SP - e12059 EP - e12059 JF - Journal of extracellular vesicles JO - J Extracell Vesicles VL - 10 IS - 4 N2 - 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. SN - 2001-3078 UR - https://www.unboundmedicine.com/medline/citation/33664936/FO_SPR_biosensor_calibrated_with_recombinant_extracellular_vesicles_enables_specific_and_sensitive_detection_directly_in_complex_matrices_ DB - PRIME DP - Unbound Medicine ER -