Open AccessFO‐SPR biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matrices
Author(s) -
Yildizhan Yagmur,
Vajrala Venkata Suresh,
Geeurickx Edward,
Declerck Charles,
Duskunovic Nevena,
De Sutter Delphine,
Noppen Sam,
Delport Filip,
Schols Dominique,
Swinnen Johannes V.,
Eyckerman Sven,
Hendrix An,
Lammertyn Jeroen,
Spasic Dragana
Publication year - 2021
Publication title -
journal of extracellular vesicles
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.94
H-Index - 68
ISSN - 2001-3078
DOI - 10.1002/jev2.12059
Subject(s) - surface plasmon resonance , cd63 , chemistry , microvesicles , detection limit , recombinant dna , chromatography , context (archaeology) , bioassay , extracellular vesicles , microbiology and biotechnology , nanotechnology , biology , biochemistry , nanoparticle , materials science , gene , paleontology , microrna , genetics
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/ B anti‐CD81 and anti‐CD63/ B anti‐CD9), resulting in 10 3 and 10 4 times higher sensitivity than the EV concentration in human blood plasma from healthy or cancer patients, respectively. Additionally, by using anti‐CD63/ B anti‐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.