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Virus detection using filament‐coupled antibodies

Virus detection using filament‐coupled antibodies Two attractive features of ELISA are the specificity of antibody–antigen recognition and the sensitivity achieved by enzymatic amplification. This report describes the development of a non‐enzymatic molecular recognition platform adaptable to point‐of‐care clinical settings and field detection of biohazardous materials. This filament‐antibody recognition assay (FARA) is based on circumferential bands of antibody probes coupled to a 120 μm diameter polyester filament. One advantage of this design is that automated processing is achieved by sequential positioning of filament‐coupled probes through a series of 25–60 μL liquid filled microcapillary chambers. This approach was evaluated by testing for the presence of M13KO7 bacterial virus using anti‐M13KO7 IgG1 monoclonal antibody coupled to a filament. Filament motion first positioned the antibodies within a microcapillary tube containing a solution of M13KO7 virus before moving the probes through subsequent chambers, where the filament‐coupled probes were washed, exposed to a fluorescently labeled anti‐M13K07 antibody, and washed again. Filament fluorescence was then measured using a flatbed microarray scanner. The presence of virus in solution produced a characteristic increase in filament fluorescence only in regions containing coupled antibody probes. Even without the enzymatic amplification of a typical ELISA, the presence of 8.3 × 108 virus particles produced a 30‐fold increase in fluorescence over an immobilized negative control antibody. In an ELISA comparison study, the filament‐based approach had a similar lower limit of sensitivity of ∼1.7 × 107 virus particles. This platform may prove attractive for point‐of‐care settings, the detection of biohazardous materials, or other applications where sensitive, rapid, and automated molecular recognition is desired. © 2005 Wiley Periodicals, Inc. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Biotechnology and Bioengineering Wiley

Virus detection using filament‐coupled antibodies

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References (67)

Publisher
Wiley
Copyright
Copyright © 2005 Wiley Periodicals, Inc., A Wiley Company
ISSN
0006-3592
eISSN
1097-0290
DOI
10.1002/bit.20537
pmid
15895380
Publisher site
See Article on Publisher Site

Abstract

Two attractive features of ELISA are the specificity of antibody–antigen recognition and the sensitivity achieved by enzymatic amplification. This report describes the development of a non‐enzymatic molecular recognition platform adaptable to point‐of‐care clinical settings and field detection of biohazardous materials. This filament‐antibody recognition assay (FARA) is based on circumferential bands of antibody probes coupled to a 120 μm diameter polyester filament. One advantage of this design is that automated processing is achieved by sequential positioning of filament‐coupled probes through a series of 25–60 μL liquid filled microcapillary chambers. This approach was evaluated by testing for the presence of M13KO7 bacterial virus using anti‐M13KO7 IgG1 monoclonal antibody coupled to a filament. Filament motion first positioned the antibodies within a microcapillary tube containing a solution of M13KO7 virus before moving the probes through subsequent chambers, where the filament‐coupled probes were washed, exposed to a fluorescently labeled anti‐M13K07 antibody, and washed again. Filament fluorescence was then measured using a flatbed microarray scanner. The presence of virus in solution produced a characteristic increase in filament fluorescence only in regions containing coupled antibody probes. Even without the enzymatic amplification of a typical ELISA, the presence of 8.3 × 108 virus particles produced a 30‐fold increase in fluorescence over an immobilized negative control antibody. In an ELISA comparison study, the filament‐based approach had a similar lower limit of sensitivity of ∼1.7 × 107 virus particles. This platform may prove attractive for point‐of‐care settings, the detection of biohazardous materials, or other applications where sensitive, rapid, and automated molecular recognition is desired. © 2005 Wiley Periodicals, Inc.

Journal

Biotechnology and BioengineeringWiley

Published: Sep 20, 2005

Keywords: molecular recognition; virus detection; ELISA

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