Label-free detection of hypoxia-induced extracellular vesicle secretion from MCF-7 cells

Nanoscale extracellular vesicles (EVs) including exosomes (50–150 nm membrane particles) have emerged as promising cancer biomarkers due to the carried genetic information about the parental cells. However the sensitive detection of these vesicles remains a challenge. Here we present a label-free el...

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Published in:	Scientific reports Vol. 8; no. 1; pp. 9402 - 9
Main Authors:	Kilic, Tugba, Valinhas, Ana Teresa De Sousa, Wall, Ivan, Renaud, Philippe, Carrara, Sandro
Format:	Journal Article
Language:	English
Published:	London Nature Publishing Group UK 20-06-2018 Nature Publishing Group
Subjects:	13/51 14/10 631/61/350/59 631/67/2322 692/53/2421 96/1 96/10 96/21 Biosensors Breast cancer CD81 antigen Cell Hypoxia - physiology Dielectric Spectroscopy Electrochemistry Enzyme-Linked Immunosorbent Assay Exosomes Extracellular vesicles Extracellular Vesicles - metabolism Humanities and Social Sciences Humans Hypoxia Immunoassays MCF-7 Cells multidisciplinary Nanoparticles Performance assessment Science Science (multidisciplinary) Sensors Spectroscopy
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Abstract	Nanoscale extracellular vesicles (EVs) including exosomes (50–150 nm membrane particles) have emerged as promising cancer biomarkers due to the carried genetic information about the parental cells. However the sensitive detection of these vesicles remains a challenge. Here we present a label-free electrochemical sensor to measure the EVs secretion levels of hypoxic and normoxic MCF-7 cells. The sensor design includes two consecutive steps; i) Au electrode surface functionalization for anti-CD81 Antibody and ii) EVs capture. The label-free detection of EVs was done via Differential Pulse Voltammetry (DPV) and Electrochemical Impedance Spectroscopy (EIS). The working linear range for the sensor was 10 2 –10 9 EVs/ml with an LOD 77 EVs/mL and 379 EVs/ml for EIS and DPV based detection. A blood-abundant protein, RhD was used for the selectivity test. In order to assess the performance of the biosensor, the level of EVs secretion by the human breast cancer MCF-7 cell line was compared with enzyme-linked immunosorbent assays (ELISA) and Nanoparticle Tracking Analysis (NTA). Designed label-free electrochemical sensors utilized for quantification of EVs secretion enhancement due to CoCl 2 -induced hypoxia and 1.23 fold increase with respect to normoxic conditions was found.
AbstractList	Nanoscale extracellular vesicles (EVs) including exosomes (50-150 nm membrane particles) have emerged as promising cancer biomarkers due to the carried genetic information about the parental cells. However the sensitive detection of these vesicles remains a challenge. Here we present a label-free electrochemical sensor to measure the EVs secretion levels of hypoxic and normoxic MCF-7 cells. The sensor design includes two consecutive steps; i) Au electrode surface functionalization for anti-CD81 Antibody and ii) EVs capture. The label-free detection of EVs was done via Differential Pulse Voltammetry (DPV) and Electrochemical Impedance Spectroscopy (EIS). The working linear range for the sensor was 102-109 EVs/ml with an LOD 77 EVs/mL and 379 EVs/ml for EIS and DPV based detection. A blood-abundant protein, RhD was used for the selectivity test. In order to assess the performance of the biosensor, the level of EVs secretion by the human breast cancer MCF-7 cell line was compared with enzyme-linked immunosorbent assays (ELISA) and Nanoparticle Tracking Analysis (NTA). Designed label-free electrochemical sensors utilized for quantification of EVs secretion enhancement due to CoCl2-induced hypoxia and 1.23 fold increase with respect to normoxic conditions was found. Nanoscale extracellular vesicles (EVs) including exosomes (50-150 nm membrane particles) have emerged as promising cancer biomarkers due to the carried genetic information about the parental cells. However the sensitive detection of these vesicles remains a challenge. Here we present a label-free electrochemical sensor to measure the EVs secretion levels of hypoxic and normoxic MCF-7 cells. The sensor design includes two consecutive steps; i) Au electrode surface functionalization for anti-CD81 Antibody and ii) EVs capture. The label-free detection of EVs was done via Differential Pulse Voltammetry (DPV) and Electrochemical Impedance Spectroscopy (EIS). The working linear range for the sensor was 10 -10 EVs/ml with an LOD 77 EVs/mL and 379 EVs/ml for EIS and DPV based detection. A blood-abundant protein, RhD was used for the selectivity test. In order to assess the performance of the biosensor, the level of EVs secretion by the human breast cancer MCF-7 cell line was compared with enzyme-linked immunosorbent assays (ELISA) and Nanoparticle Tracking Analysis (NTA). Designed label-free electrochemical sensors utilized for quantification of EVs secretion enhancement due to CoCl -induced hypoxia and 1.23 fold increase with respect to normoxic conditions was found. Nanoscale extracellular vesicles (EVs) including exosomes (50–150 nm membrane particles) have emerged as promising cancer biomarkers due to the carried genetic information about the parental cells. However the sensitive detection of these vesicles remains a challenge. Here we present a label-free electrochemical sensor to measure the EVs secretion levels of hypoxic and normoxic MCF-7 cells. The sensor design includes two consecutive steps; i) Au electrode surface functionalization for anti-CD81 Antibody and ii) EVs capture. The label-free detection of EVs was done via Differential Pulse Voltammetry (DPV) and Electrochemical Impedance Spectroscopy (EIS). The working linear range for the sensor was 10 2 –10 9 EVs/ml with an LOD 77 EVs/mL and 379 EVs/ml for EIS and DPV based detection. A blood-abundant protein, RhD was used for the selectivity test. In order to assess the performance of the biosensor, the level of EVs secretion by the human breast cancer MCF-7 cell line was compared with enzyme-linked immunosorbent assays (ELISA) and Nanoparticle Tracking Analysis (NTA). Designed label-free electrochemical sensors utilized for quantification of EVs secretion enhancement due to CoCl 2 -induced hypoxia and 1.23 fold increase with respect to normoxic conditions was found. Nanoscale extracellular vesicles (EVs) including exosomes (50–150 nm membrane particles) have emerged as promising cancer biomarkers due to the carried genetic information about the parental cells. However the sensitive detection of these vesicles remains a challenge. Here we present a label-free electrochemical sensor to measure the EVs secretion levels of hypoxic and normoxic MCF-7 cells. The sensor design includes two consecutive steps; i) Au electrode surface functionalization for anti-CD81 Antibody and ii) EVs capture. The label-free detection of EVs was done via Differential Pulse Voltammetry (DPV) and Electrochemical Impedance Spectroscopy (EIS). The working linear range for the sensor was 10 2 –10 9 EVs/ml with an LOD 77 EVs/mL and 379 EVs/ml for EIS and DPV based detection. A blood-abundant protein, RhD was used for the selectivity test. In order to assess the performance of the biosensor, the level of EVs secretion by the human breast cancer MCF-7 cell line was compared with enzyme-linked immunosorbent assays (ELISA) and Nanoparticle Tracking Analysis (NTA). Designed label-free electrochemical sensors utilized for quantification of EVs secretion enhancement due to CoCl 2 -induced hypoxia and 1.23 fold increase with respect to normoxic conditions was found.
ArticleNumber	9402
Author	Carrara, Sandro Valinhas, Ana Teresa De Sousa Wall, Ivan Renaud, Philippe Kilic, Tugba
Author_xml	– sequence: 1 givenname: Tugba orcidid: 0000-0001-8600-0367 surname: Kilic fullname: Kilic, Tugba email: tugba.kilic@epfl.ch organization: Swiss Federal Institute of Technology Lausanne, EPFL, Integrated Systems Laboratory (LSI), Swiss Federal Institute of Technology Lausanne, EPFL, Microsystems Laboratory 4 (LMIS4) – sequence: 2 givenname: Ana Teresa De Sousa surname: Valinhas fullname: Valinhas, Ana Teresa De Sousa organization: Swiss Federal Institute of Technology Lausanne, EPFL, Integrated Systems Laboratory (LSI), Swiss Federal Institute of Technology Lausanne, EPFL, Microsystems Laboratory 4 (LMIS4), Department of Biochemical Engineering, University College London, Bernard Katz Building – sequence: 3 givenname: Ivan surname: Wall fullname: Wall, Ivan organization: Department of Biochemical Engineering, University College London, Bernard Katz Building – sequence: 4 givenname: Philippe surname: Renaud fullname: Renaud, Philippe organization: Swiss Federal Institute of Technology Lausanne, EPFL, Microsystems Laboratory 4 (LMIS4) – sequence: 5 givenname: Sandro surname: Carrara fullname: Carrara, Sandro organization: Swiss Federal Institute of Technology Lausanne, EPFL, Integrated Systems Laboratory (LSI)
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Snippet	Nanoscale extracellular vesicles (EVs) including exosomes (50–150 nm membrane particles) have emerged as promising cancer biomarkers due to the carried genetic... Nanoscale extracellular vesicles (EVs) including exosomes (50-150 nm membrane particles) have emerged as promising cancer biomarkers due to the carried genetic... Nanoscale extracellular vesicles (EVs) including exosomes (50–150 nm membrane particles) have emerged as promising cancer biomarkers due to the carried genetic...
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SubjectTerms	13/51 14/10 631/61/350/59 631/67/2322 692/53/2421 96/1 96/10 96/21 Biosensors Breast cancer CD81 antigen Cell Hypoxia - physiology Dielectric Spectroscopy Electrochemistry Enzyme-Linked Immunosorbent Assay Exosomes Extracellular vesicles Extracellular Vesicles - metabolism Humanities and Social Sciences Humans Hypoxia Immunoassays MCF-7 Cells multidisciplinary Nanoparticles Performance assessment Science Science (multidisciplinary) Sensors Spectroscopy
Title	Label-free detection of hypoxia-induced extracellular vesicle secretion from MCF-7 cells
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