Paper-based microfluidics is a new technology which utilizes paper as the fabrication substrate to pattern microstructures and has been shown to be simple, inexpensive, and an easy-to-use diagnostic platform. Nitrocellulose (NC) membranes have seen limited use but their characteristic properties should be amenable to microfluidic-based applications even more than paper or other soft-lithographic techniques. Herein, we describe a novel paper-based microfluidic device (MD) assay utilizing an NC membrane to detect immunoglobulin G (IgG) antibodies via a colorimetric analysis. The MDs consisted of layered polyethylene terephthalate and pressure-sensitive adhesives and an NC channel. The biotin labeled IgG antibody was spotted and dried on the NC channel before wicking a series of solutions through it, including a Tris-tween wash and varying concentrations of alkaline phosphatase conjugated to streptavidin (ALP-Strep). Para-nitrophenyl phosphate (pNPP) was spotted at the site of the antibody following these solutions, which results in a vibrant yellow color. The reaction proceeds for 10 minutes before applying the pNPP stop solution. The device was then dried, scanned, and analyzed yielding a linear range of yellow color intensities vs. ALP-Strep concentrations.

Development of paper-based microfluidic enzyme-linked immunosorbent assays (ELISAs)

248th National Meeting (2014)

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