A Novel Microfluidic Point-of-Care Biosensor System on Printed Circuit Board for Cytokine Detection
Point of Care (PoC) diagnostics have been the subject of considerable research over the last few decades driven by the pressure to detect diseases quickly and effectively and reduce healthcare costs. Herein, we demonstrate a novel, fully integrated, microfluidic amperometric enzyme-linked immunosorb...
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| Published in | Sensors (Basel, Switzerland) Vol. 18; no. 11; p. 4011 |
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| Main Authors | , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Switzerland
MDPI
17.11.2018
MDPI AG |
| Subjects | |
| Online Access | Get full text |
| ISSN | 1424-8220 1424-8220 |
| DOI | 10.3390/s18114011 |
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| Abstract | Point of Care (PoC) diagnostics have been the subject of considerable research over the last few decades driven by the pressure to detect diseases quickly and effectively and reduce healthcare costs. Herein, we demonstrate a novel, fully integrated, microfluidic amperometric enzyme-linked immunosorbent assay (ELISA) prototype using a commercial interferon gamma release assay (IGRA) as a model antibody binding system. Microfluidic assay chemistry was engineered to take place on Au-plated electrodes within an assay cell on a printed circuit board (PCB)-based biosensor system. The assay cell is linked to an electrochemical reporter cell comprising microfluidic architecture, Au working and counter electrodes and a Ag/AgCl reference electrode, all manufactured exclusively via standard commercial PCB fabrication processes. Assay chemistry has been optimised for microfluidic diffusion kinetics to function under continual flow. We characterised the electrode integrity of the developed platforms with reference to biological sampling and buffer composition and subsequently we demonstrated concentration-dependent measurements of H2O2 depletion as resolved by existing FDA-validated ELISA kits. Finally, we validated the assay technology in both buffer and serum and demonstrate limits of detection comparable to high-end commercial systems with the addition of full microfluidic assay architecture capable of returning diagnostic analyses in approximately eight minutes. |
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| AbstractList | Point of Care (PoC) diagnostics have been the subject of considerable research over the last few decades driven by the pressure to detect diseases quickly and effectively and reduce healthcare costs. Herein, we demonstrate a novel, fully integrated, microfluidic amperometric enzyme-linked immunosorbent assay (ELISA) prototype using a commercial interferon gamma release assay (IGRA) as a model antibody binding system. Microfluidic assay chemistry was engineered to take place on Au-plated electrodes within an assay cell on a printed circuit board (PCB)-based biosensor system. The assay cell is linked to an electrochemical reporter cell comprising microfluidic architecture, Au working and counter electrodes and a Ag/AgCl reference electrode, all manufactured exclusively via standard commercial PCB fabrication processes. Assay chemistry has been optimised for microfluidic diffusion kinetics to function under continual flow. We characterised the electrode integrity of the developed platforms with reference to biological sampling and buffer composition and subsequently we demonstrated concentration-dependent measurements of H2O2 depletion as resolved by existing FDA-validated ELISA kits. Finally, we validated the assay technology in both buffer and serum and demonstrate limits of detection comparable to high-end commercial systems with the addition of full microfluidic assay architecture capable of returning diagnostic analyses in approximately eight minutes. Point of Care (PoC) diagnostics have been the subject of considerable research over the last few decades driven by the pressure to detect diseases quickly and effectively and reduce healthcare costs. Herein, we demonstrate a novel, fully integrated, microfluidic amperometric enzyme-linked immunosorbent assay (ELISA) prototype using a commercial interferon gamma release assay (IGRA) as a model antibody binding system. Microfluidic assay chemistry was engineered to take place on Au-plated electrodes within an assay cell on a printed circuit board (PCB)-based biosensor system. The assay cell is linked to an electrochemical reporter cell comprising microfluidic architecture, Au working and counter electrodes and a Ag/AgCl reference electrode, all manufactured exclusively via standard commercial PCB fabrication processes. Assay chemistry has been optimised for microfluidic diffusion kinetics to function under continual flow. We characterised the electrode integrity of the developed platforms with reference to biological sampling and buffer composition and subsequently we demonstrated concentration-dependent measurements of H 2 O 2 depletion as resolved by existing FDA-validated ELISA kits. Finally, we validated the assay technology in both buffer and serum and demonstrate limits of detection comparable to high-end commercial systems with the addition of full microfluidic assay architecture capable of returning diagnostic analyses in approximately eight minutes. Point of Care (PoC) diagnostics have been the subject of considerable research over the last few decades driven by the pressure to detect diseases quickly and effectively and reduce healthcare costs. Herein, we demonstrate a novel, fully integrated, microfluidic amperometric enzyme-linked immunosorbent assay (ELISA) prototype using a commercial interferon gamma release assay (IGRA) as a model antibody binding system. Microfluidic assay chemistry was engineered to take place on Au-plated electrodes within an assay cell on a printed circuit board (PCB)-based biosensor system. The assay cell is linked to an electrochemical reporter cell comprising microfluidic architecture, Au working and counter electrodes and a Ag/AgCl reference electrode, all manufactured exclusively via standard commercial PCB fabrication processes. Assay chemistry has been optimised for microfluidic diffusion kinetics to function under continual flow. We characterised the electrode integrity of the developed platforms with reference to biological sampling and buffer composition and subsequently we demonstrated concentration-dependent measurements of H₂O₂ depletion as resolved by existing FDA-validated ELISA kits. Finally, we validated the assay technology in both buffer and serum and demonstrate limits of detection comparable to high-end commercial systems with the addition of full microfluidic assay architecture capable of returning diagnostic analyses in approximately eight minutes.Point of Care (PoC) diagnostics have been the subject of considerable research over the last few decades driven by the pressure to detect diseases quickly and effectively and reduce healthcare costs. Herein, we demonstrate a novel, fully integrated, microfluidic amperometric enzyme-linked immunosorbent assay (ELISA) prototype using a commercial interferon gamma release assay (IGRA) as a model antibody binding system. Microfluidic assay chemistry was engineered to take place on Au-plated electrodes within an assay cell on a printed circuit board (PCB)-based biosensor system. The assay cell is linked to an electrochemical reporter cell comprising microfluidic architecture, Au working and counter electrodes and a Ag/AgCl reference electrode, all manufactured exclusively via standard commercial PCB fabrication processes. Assay chemistry has been optimised for microfluidic diffusion kinetics to function under continual flow. We characterised the electrode integrity of the developed platforms with reference to biological sampling and buffer composition and subsequently we demonstrated concentration-dependent measurements of H₂O₂ depletion as resolved by existing FDA-validated ELISA kits. Finally, we validated the assay technology in both buffer and serum and demonstrate limits of detection comparable to high-end commercial systems with the addition of full microfluidic assay architecture capable of returning diagnostic analyses in approximately eight minutes. Point of Care (PoC) diagnostics have been the subject of considerable research over the last few decades driven by the pressure to detect diseases quickly and effectively and reduce healthcare costs. Herein, we demonstrate a novel, fully integrated, microfluidic amperometric enzyme-linked immunosorbent assay (ELISA) prototype using a commercial interferon gamma release assay (IGRA) as a model antibody binding system. Microfluidic assay chemistry was engineered to take place on Au-plated electrodes within an assay cell on a printed circuit board (PCB)-based biosensor system. The assay cell is linked to an electrochemical reporter cell comprising microfluidic architecture, Au working and counter electrodes and a Ag/AgCl reference electrode, all manufactured exclusively via standard commercial PCB fabrication processes. Assay chemistry has been optimised for microfluidic diffusion kinetics to function under continual flow. We characterised the electrode integrity of the developed platforms with reference to biological sampling and buffer composition and subsequently we demonstrated concentration-dependent measurements of H₂O₂ depletion as resolved by existing FDA-validated ELISA kits. Finally, we validated the assay technology in both buffer and serum and demonstrate limits of detection comparable to high-end commercial systems with the addition of full microfluidic assay architecture capable of returning diagnostic analyses in approximately eight minutes. |
| Author | Vasilakis, Nikolaos Pantelidis, Panagiotis Kelleher, Peter Papadimitriou, Konstantinos Morgan, Hywel Prodromakis, Themistoklis Evans, Daniel |
| AuthorAffiliation | 1 Nanoelectronics & Nanotechnology Research Group, Electronics and Computer Science, University of Southampton, Southampton SO17 1BJ, UK; k.papadimitriou@ucl.ac.uk (K.I.P.); n.vasilakis@soton.ac.uk (N.V.); hm@ecs.soton.ac.uk (H.M.) 2 Centre for Immunology and Vaccinology, Division of Infectious Diseases, Department of Medicine, Imperial College London, London SW10 9NH, UK; panagiotis.pantelidis@nhs.net (P.P.); p.kelleher@imperial.ac.uk (P.K.) 4 Institute for Life Sciences, University of Southampton, Southampton SO17 1BJ, UK 5 Zepler Institute for Photonics and Nanoelectronics, University of Southampton, Southampton SO17 1BJ, UK; t.prodromakis@soton.ac.uk 3 Infection and Immunity, North West London Pathology, Imperial College NHS Trust, Charing Cross Hospital, London W6 8RF, UK |
| AuthorAffiliation_xml | – name: 1 Nanoelectronics & Nanotechnology Research Group, Electronics and Computer Science, University of Southampton, Southampton SO17 1BJ, UK; k.papadimitriou@ucl.ac.uk (K.I.P.); n.vasilakis@soton.ac.uk (N.V.); hm@ecs.soton.ac.uk (H.M.) – name: 2 Centre for Immunology and Vaccinology, Division of Infectious Diseases, Department of Medicine, Imperial College London, London SW10 9NH, UK; panagiotis.pantelidis@nhs.net (P.P.); p.kelleher@imperial.ac.uk (P.K.) – name: 3 Infection and Immunity, North West London Pathology, Imperial College NHS Trust, Charing Cross Hospital, London W6 8RF, UK – name: 5 Zepler Institute for Photonics and Nanoelectronics, University of Southampton, Southampton SO17 1BJ, UK; t.prodromakis@soton.ac.uk – name: 4 Institute for Life Sciences, University of Southampton, Southampton SO17 1BJ, UK |
| Author_xml | – sequence: 1 givenname: Daniel surname: Evans fullname: Evans, Daniel – sequence: 2 givenname: Konstantinos orcidid: 0000-0002-0995-4834 surname: Papadimitriou fullname: Papadimitriou, Konstantinos – sequence: 3 givenname: Nikolaos surname: Vasilakis fullname: Vasilakis, Nikolaos – sequence: 4 givenname: Panagiotis surname: Pantelidis fullname: Pantelidis, Panagiotis – sequence: 5 givenname: Peter surname: Kelleher fullname: Kelleher, Peter – sequence: 6 givenname: Hywel surname: Morgan fullname: Morgan, Hywel – sequence: 7 givenname: Themistoklis surname: Prodromakis fullname: Prodromakis, Themistoklis |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/30453609$$D View this record in MEDLINE/PubMed |
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| Keywords | point-of-care diagnostics eELISA lab-on-PCB PCB biosensors cytokine detection microfluidics |
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| Title | A Novel Microfluidic Point-of-Care Biosensor System on Printed Circuit Board for Cytokine Detection |
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