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Development of a microfluidic‐based assay on a novel nitrocellulose platform
Authors:Mary Arrastia  Ani Avoundjian  Paul Said Ehrlich  Micah Eropkin  Leanna Levine  Frank A. Gomez
Affiliation:1. Department of Chemistry and Biochemistry, California State University, Los Angeles, CA, USA;2. ALine Inc, Rancho Dominguez, CA, USA
Abstract:A novel microfluidic paper‐based analytical device (μPAD) utilizing a nitrocellulose (NC) membrane to detect IgG antibodies through a colorimetric analysis is described. The μPAD was constructed using layered polyethylene terephthalate (PET) and pressure‐sensitive adhesives (PSA). The biotin labeled Goat Anti‐Mouse IgG antibody was spotted and dried on the NC channel prior to subjecting it to a series of wash solutions (Tris‐tween), increasing concentrations of alkaline phosphatase conjugated to streptavidin (Strep‐ALP), and para‐nitrophenyl phosphate (p‐NPP) realizing a vibrant yellow color. The reaction proceeds for 10 min before applying the p‐NPP stop solution. The device was then dried, scanned, and analyzed yielding a linear range of inverse yellow color intensities versus Strep‐ALP concentrations. The development of this simple μPAD should further facilitate the use of NC in colorimetric assays to detect and quantitate antibodies.
Keywords:ELISA  Microfluidic paper‐based analytical device  Microfluidics
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