Presentation Title

Thread/Paper-Based Enzyme-Linked Immunosorbent Assay (ELISA)

Faculty Mentor

Frank A. Gomez

Start Date

18-11-2017 10:00 AM

End Date

18-11-2017 10:15 AM

Location

9-263

Session

Physical Sciences 3

Type of Presentation

Oral Talk

Subject Area

physical_mathematical_sciences

Abstract

We demonstrate the use of a novel microfluidic thread/paper-based analytical device (µTPAD) for the detection of biotinylated goat anti-mouse IgG (system one) and rabbit IgG (system two) antibodies via colorimetric analysis. Fabrication of the µTPAD consist of six pieces of nylon thread, three pieces of flannel, and three pieces of circular paper chromatography analysis sites. For system one, nitrocellulose (NC) was spotted on the detection sites. Biotinylated goat anti-mouse IgG was spotted and dried on the analysis sites, prior to subjecting it to a series of Tris-Tween washes, increasing streptavidin-alkaline phosphatase (Strep-ALP) concentrations, p-nitrophenyl phosphate (p-NPP), and stop p-NPP solution, realizing a yellow color at the detection sites. The detection sites were then scanned and reported as inverse yellow intensity versus concentration of Strep-ALP. The yellow color intensity increases as Strep-ALP concentration increases. For system two, primary rabbit IgG antibody was spotted and dried on the analysis sites, followed by a solution of Tween-BSA, ALP-conjugated secondary antibody, a series of PBS washes, and a solution of the colorimetric substrate for ALP, realizing a purple color at the detection sites. The platforms were scanned and analyzed yielding a correlation between the purple color produced, which was analyzed as inverse blue intensity, and the concentration of rabbit IgG. The purple color increased as the concentration of the primary antibody increased. The design of the µTPAD enables triplicate data collection. The device is low-cost, easy-to-use, requires a minimal amount of material, and facilitates an ELISA.

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Nov 18th, 10:00 AM Nov 18th, 10:15 AM

Thread/Paper-Based Enzyme-Linked Immunosorbent Assay (ELISA)

9-263

We demonstrate the use of a novel microfluidic thread/paper-based analytical device (µTPAD) for the detection of biotinylated goat anti-mouse IgG (system one) and rabbit IgG (system two) antibodies via colorimetric analysis. Fabrication of the µTPAD consist of six pieces of nylon thread, three pieces of flannel, and three pieces of circular paper chromatography analysis sites. For system one, nitrocellulose (NC) was spotted on the detection sites. Biotinylated goat anti-mouse IgG was spotted and dried on the analysis sites, prior to subjecting it to a series of Tris-Tween washes, increasing streptavidin-alkaline phosphatase (Strep-ALP) concentrations, p-nitrophenyl phosphate (p-NPP), and stop p-NPP solution, realizing a yellow color at the detection sites. The detection sites were then scanned and reported as inverse yellow intensity versus concentration of Strep-ALP. The yellow color intensity increases as Strep-ALP concentration increases. For system two, primary rabbit IgG antibody was spotted and dried on the analysis sites, followed by a solution of Tween-BSA, ALP-conjugated secondary antibody, a series of PBS washes, and a solution of the colorimetric substrate for ALP, realizing a purple color at the detection sites. The platforms were scanned and analyzed yielding a correlation between the purple color produced, which was analyzed as inverse blue intensity, and the concentration of rabbit IgG. The purple color increased as the concentration of the primary antibody increased. The design of the µTPAD enables triplicate data collection. The device is low-cost, easy-to-use, requires a minimal amount of material, and facilitates an ELISA.