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Highly sensitive immunoassay based on controlled rehydration of patterned reagents in a 2-dimensional paper network.
Anal Chem. 2014 Jul 01; 86(13):6447-53.AC

Abstract

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.

Authors+Show Affiliations

University of Washington , Department of Bioengineering, Box 355061, Seattle, Washington 98195, United States.No affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

24882058

Citation

Fridley, Gina E., et al. "Highly Sensitive Immunoassay Based On Controlled Rehydration of Patterned Reagents in a 2-dimensional Paper Network." Analytical Chemistry, vol. 86, no. 13, 2014, pp. 6447-53.
Fridley GE, Le H, Yager P. Highly sensitive immunoassay based on controlled rehydration of patterned reagents in a 2-dimensional paper network. Anal Chem. 2014;86(13):6447-53.
Fridley, G. E., Le, H., & Yager, P. (2014). Highly sensitive immunoassay based on controlled rehydration of patterned reagents in a 2-dimensional paper network. Analytical Chemistry, 86(13), 6447-53. https://doi.org/10.1021/ac500872j
Fridley GE, Le H, Yager P. Highly Sensitive Immunoassay Based On Controlled Rehydration of Patterned Reagents in a 2-dimensional Paper Network. Anal Chem. 2014 Jul 1;86(13):6447-53. PubMed PMID: 24882058.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Highly sensitive immunoassay based on controlled rehydration of patterned reagents in a 2-dimensional paper network. AU - Fridley,Gina E, AU - Le,Huy, AU - Yager,Paul, Y1 - 2014/06/17/ PY - 2014/6/3/entrez PY - 2014/6/3/pubmed PY - 2015/8/19/medline SP - 6447 EP - 53 JF - Analytical chemistry JO - Anal. Chem. VL - 86 IS - 13 N2 - 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. SN - 1520-6882 UR - https://www.unboundmedicine.com/medline/citation/24882058/Highly_sensitive_immunoassay_based_on_controlled_rehydration_of_patterned_reagents_in_a_2_dimensional_paper_network_ L2 - https://dx.doi.org/10.1021/ac500872j DB - PRIME DP - Unbound Medicine ER -