2015
DOI: 10.1007/s00216-015-9052-0
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Immobilizing affinity proteins to nitrocellulose: a toolbox for paper-based assay developers

Abstract: To enable enhanced paper-based diagnostics with improved detection capabilities, new methods are needed to immobilize affinity reagents to porous substrates, especially for capture molecules other than IgG. To this end, we have developed and characterized three novel methods for immobilizing protein-based affinity reagents to nitrocellulose membranes. We have demonstrated these methods using recombinant affinity proteins for the influenza surface protein hemagglutinin, leveraging the customizability of these r… Show more

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Cited by 78 publications
(68 citation statements)
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“…We tested the utility of Tri-HSB for detecting HA in a paper-based assay similar to what might be used for diagnosis of infectious diseases at point-of-care sites or in home-testing kits 25 . As shown in Figure 4a, Tri-HSB is effective both for capturing and detecting HA in a lateral flow assay, with a very clear band on the nitrocellulose strip and little or no background using either gold nanoparticles or diaminobenzidine (DAB) precipitation by conjugated HRP.…”
mentioning
confidence: 99%
“…We tested the utility of Tri-HSB for detecting HA in a paper-based assay similar to what might be used for diagnosis of infectious diseases at point-of-care sites or in home-testing kits 25 . As shown in Figure 4a, Tri-HSB is effective both for capturing and detecting HA in a lateral flow assay, with a very clear band on the nitrocellulose strip and little or no background using either gold nanoparticles or diaminobenzidine (DAB) precipitation by conjugated HRP.…”
mentioning
confidence: 99%
“…A tailored antibody can be further developed into a paper-based diagnostics or biosensors [13]. Recently, four new methods to immobilize antibody on the nitrocellulose were introduced to support the rapid test developer [14].…”
Section: Discussionmentioning
confidence: 99%
“…These immobilization strategies have been already employed in the fields of ELISA and lateral flow assay, which were exclusively based on passive adsorption of antibody. [7,44]…”
Section: Antibody Immobilization Chemistriesmentioning
confidence: 99%