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Highly Sensitive Immunoassay Based on Controlled Rehydration of Patterned Reagents in a 2-Dimensional Paper Network

[Image: see text] We have demonstrated a multistep 2-dimensional paper network immunoassay based on controlled rehydration of patterned, dried reagents. Previous work has shown that signal enhancement improves the limit of detection in 2-dimensional paper network assays, but until now, reagents have...

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Autores principales: Fridley, Gina E., Le, Huy, Yager, Paul
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2014
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4082385/
https://www.ncbi.nlm.nih.gov/pubmed/24882058
http://dx.doi.org/10.1021/ac500872j
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author Fridley, Gina E.
Le, Huy
Yager, Paul
author_facet Fridley, Gina E.
Le, Huy
Yager, Paul
author_sort Fridley, Gina E.
collection PubMed
description [Image: see text] We have demonstrated a multistep 2-dimensional paper network immunoassay based on controlled rehydration of patterned, dried reagents. Previous work has shown that signal enhancement improves the limit of detection in 2-dimensional paper network assays, but until now, reagents have only been included as wet or dried in separate conjugate pads placed at the upstream end of the assay device. Wet reagents are not ideal for point-of-care because they must be refrigerated and typically limit automation and require more user steps. Conjugate pads allow drying but do not offer any control of the reagent distribution upon rehydration and can be a source of error when pads do not contact the assay membrane uniformly. Furthermore, each reagent is dried on a separate pad, increasing the fabrication complexity when implementing multistep assays that require several different reagents. Conversely, our novel method allows for consistent, controlled rehydration from patterned reagent storage depots directly within the paper membrane. In this assay demonstration, four separate reagents were patterned in different regions of the assay device: a gold-antibody conjugate used for antigen detection and three different signal enhancement components that must not be mixed until immediately before use. To show the viability of patterning and drying reagents directly onto a paper device for dry reagent storage and subsequent controlled release, we tested this device with the malaria antigen Plasmodium falciparum histidine-rich protein 2 (PfHRP2) as an example of target analyte. In this demonstration, the signal enhancement step increases the visible signal by roughly 3-fold and decreases the analytical limit of detection by 2.75-fold.
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spelling pubmed-40823852015-05-31 Highly Sensitive Immunoassay Based on Controlled Rehydration of Patterned Reagents in a 2-Dimensional Paper Network Fridley, Gina E. Le, Huy Yager, Paul Anal Chem [Image: see text] We have demonstrated a multistep 2-dimensional paper network immunoassay based on controlled rehydration of patterned, dried reagents. Previous work has shown that signal enhancement improves the limit of detection in 2-dimensional paper network assays, but until now, reagents have only been included as wet or dried in separate conjugate pads placed at the upstream end of the assay device. Wet reagents are not ideal for point-of-care because they must be refrigerated and typically limit automation and require more user steps. Conjugate pads allow drying but do not offer any control of the reagent distribution upon rehydration and can be a source of error when pads do not contact the assay membrane uniformly. Furthermore, each reagent is dried on a separate pad, increasing the fabrication complexity when implementing multistep assays that require several different reagents. Conversely, our novel method allows for consistent, controlled rehydration from patterned reagent storage depots directly within the paper membrane. In this assay demonstration, four separate reagents were patterned in different regions of the assay device: a gold-antibody conjugate used for antigen detection and three different signal enhancement components that must not be mixed until immediately before use. To show the viability of patterning and drying reagents directly onto a paper device for dry reagent storage and subsequent controlled release, we tested this device with the malaria antigen Plasmodium falciparum histidine-rich protein 2 (PfHRP2) as an example of target analyte. In this demonstration, the signal enhancement step increases the visible signal by roughly 3-fold and decreases the analytical limit of detection by 2.75-fold. American Chemical Society 2014-05-31 2014-07-01 /pmc/articles/PMC4082385/ /pubmed/24882058 http://dx.doi.org/10.1021/ac500872j Text en Copyright © 2014 American Chemical Society Terms of Use (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html)
spellingShingle Fridley, Gina E.
Le, Huy
Yager, Paul
Highly Sensitive Immunoassay Based on Controlled Rehydration of Patterned Reagents in a 2-Dimensional Paper Network
title Highly Sensitive Immunoassay Based on Controlled Rehydration of Patterned Reagents in a 2-Dimensional Paper Network
title_full Highly Sensitive Immunoassay Based on Controlled Rehydration of Patterned Reagents in a 2-Dimensional Paper Network
title_fullStr Highly Sensitive Immunoassay Based on Controlled Rehydration of Patterned Reagents in a 2-Dimensional Paper Network
title_full_unstemmed Highly Sensitive Immunoassay Based on Controlled Rehydration of Patterned Reagents in a 2-Dimensional Paper Network
title_short Highly Sensitive Immunoassay Based on Controlled Rehydration of Patterned Reagents in a 2-Dimensional Paper Network
title_sort highly sensitive immunoassay based on controlled rehydration of patterned reagents in a 2-dimensional paper network
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4082385/
https://www.ncbi.nlm.nih.gov/pubmed/24882058
http://dx.doi.org/10.1021/ac500872j
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