title: Development of a microfluidic-based assay on a novel nitrocellulose platform.
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.
authors.affiliation: Department of Chemistry and Biochemistry, California State University, Los Angeles, CA, USA.
authors.forename: Mary
authors.initials: M
authors.lastname: Arrastia
authors.forename: Ani
authors.initials: A
authors.lastname: Avoundjian
authors.forename: Paul Said
authors.initials: PS
authors.lastname: Ehrlich
authors.forename: Micah
authors.initials: M
authors.lastname: Eropkin
authors.forename: Leanna
authors.initials: L
authors.lastname: Levine
authors.forename: Frank A
authors.initials: FA
authors.lastname: Gomez
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry; D003101:Collodion / Q000737:chemistry*; D003124:Colorimetry / Q000295:instrumentation*; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation*; D004867:Equipment Design; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*
chemical_list: D000906:Antibodies; D003101:Collodion
keywords: ELISA; Microfluidic paper-based analytical device; Microfluidics