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Rapid Multianalyte Microfluidic Homogeneous Immunoassay on Electrokinetically Driven Beads

The simplicity of homogeneous immunoassays makes them suitable for diagnostics of acute conditions. Indeed, the absence of washing steps reduces the binding reaction duration and favors a rapid and compact device, a critical asset for patients experiencing life-threatening diseases. In order to maxi...

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Detalles Bibliográficos
Autores principales: Thiriet, Pierre-Emmanuel, Medagoda, Danashi, Porro, Gloria, Guiducci, Carlotta
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7766682/
https://www.ncbi.nlm.nih.gov/pubmed/33371213
http://dx.doi.org/10.3390/bios10120212
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author Thiriet, Pierre-Emmanuel
Medagoda, Danashi
Porro, Gloria
Guiducci, Carlotta
author_facet Thiriet, Pierre-Emmanuel
Medagoda, Danashi
Porro, Gloria
Guiducci, Carlotta
author_sort Thiriet, Pierre-Emmanuel
collection PubMed
description The simplicity of homogeneous immunoassays makes them suitable for diagnostics of acute conditions. Indeed, the absence of washing steps reduces the binding reaction duration and favors a rapid and compact device, a critical asset for patients experiencing life-threatening diseases. In order to maximize analytical performance, standard systems employed in clinical laboratories rely largely on the use of high surface-to-volume ratio suspended moieties, such as microbeads, which provide at the same time a fast and efficient collection of analytes from the sample and controlled aggregation of collected material for improved readout. Here, we introduce an integrated microfluidic system that can perform analyte detection on antibody-decorated beads and their accumulation in confined regions within 15 min. We employed the system to the concomitant analysis of clinical concentrations of Neutrophil Gelatinase-Associated Lipocalin (NGAL) and Cystatin C in serum, two acute kidney injury (AKI) biomarkers. To this end, high-aspect-ratio, three-dimensional electrodes were integrated within a microfluidic channel to impart a controlled trajectory to antibody-decorated microbeads through the application of dielectrophoretic (DEP) forces. Beads were efficiently retained against the fluid flow of reagents, granting an efficient on-chip analyte-to-bead binding. Electrokinetic forces specific to the beads’ size were generated in the same channel, leading differently decorated beads to different readout regions of the chip. Therefore, this microfluidic multianalyte immunoassay was demonstrated as a powerful tool for the rapid detection of acute life-threatening conditions.
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spelling pubmed-77666822020-12-28 Rapid Multianalyte Microfluidic Homogeneous Immunoassay on Electrokinetically Driven Beads Thiriet, Pierre-Emmanuel Medagoda, Danashi Porro, Gloria Guiducci, Carlotta Biosensors (Basel) Article The simplicity of homogeneous immunoassays makes them suitable for diagnostics of acute conditions. Indeed, the absence of washing steps reduces the binding reaction duration and favors a rapid and compact device, a critical asset for patients experiencing life-threatening diseases. In order to maximize analytical performance, standard systems employed in clinical laboratories rely largely on the use of high surface-to-volume ratio suspended moieties, such as microbeads, which provide at the same time a fast and efficient collection of analytes from the sample and controlled aggregation of collected material for improved readout. Here, we introduce an integrated microfluidic system that can perform analyte detection on antibody-decorated beads and their accumulation in confined regions within 15 min. We employed the system to the concomitant analysis of clinical concentrations of Neutrophil Gelatinase-Associated Lipocalin (NGAL) and Cystatin C in serum, two acute kidney injury (AKI) biomarkers. To this end, high-aspect-ratio, three-dimensional electrodes were integrated within a microfluidic channel to impart a controlled trajectory to antibody-decorated microbeads through the application of dielectrophoretic (DEP) forces. Beads were efficiently retained against the fluid flow of reagents, granting an efficient on-chip analyte-to-bead binding. Electrokinetic forces specific to the beads’ size were generated in the same channel, leading differently decorated beads to different readout regions of the chip. Therefore, this microfluidic multianalyte immunoassay was demonstrated as a powerful tool for the rapid detection of acute life-threatening conditions. MDPI 2020-12-21 /pmc/articles/PMC7766682/ /pubmed/33371213 http://dx.doi.org/10.3390/bios10120212 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Thiriet, Pierre-Emmanuel
Medagoda, Danashi
Porro, Gloria
Guiducci, Carlotta
Rapid Multianalyte Microfluidic Homogeneous Immunoassay on Electrokinetically Driven Beads
title Rapid Multianalyte Microfluidic Homogeneous Immunoassay on Electrokinetically Driven Beads
title_full Rapid Multianalyte Microfluidic Homogeneous Immunoassay on Electrokinetically Driven Beads
title_fullStr Rapid Multianalyte Microfluidic Homogeneous Immunoassay on Electrokinetically Driven Beads
title_full_unstemmed Rapid Multianalyte Microfluidic Homogeneous Immunoassay on Electrokinetically Driven Beads
title_short Rapid Multianalyte Microfluidic Homogeneous Immunoassay on Electrokinetically Driven Beads
title_sort rapid multianalyte microfluidic homogeneous immunoassay on electrokinetically driven beads
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7766682/
https://www.ncbi.nlm.nih.gov/pubmed/33371213
http://dx.doi.org/10.3390/bios10120212
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