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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical
Society
2014
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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. |
format | Online Article Text |
id | pubmed-4082385 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | American Chemical
Society |
record_format | MEDLINE/PubMed |
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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