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The thermally responsive hydrogel N,N'-methylenebisacrylamide-cross-linked poly(N-isopropylacrylamide) (PNIPAM) was developed and evaluated as a reagent storage and delivery system for microfluidic paper-based analytical devices (microPADs). PNIPAM was shown to successfully deliver multiple solutions to microPADs in specific sequences or simultaneously in laminar-flow configuration and was found to be suitable for delivering four classes of reagents to the devices: Small molecules, enzymes, antibodies and DNA. PNIPAM was also able to successfully deliver a series of standard glucose solutions to microPADs equipped to perform a colorimetric glucose assay. The results of these tests were used to produce an external calibration curve, which in turn was used to determine the concentration of glucose in sample solutions. Finally, PNIPAM was used to store the enzyme horseradish peroxidase for 35 days under ambient conditions with no significant loss of activity. The combination of PNIPAM and microPADs may allow for more complex assays to be performed on paper-based devices, facilitate the preparation of external calibration curves in the field, and extend the shelf life of microPADs by stabilizing reagents in an  easy-to-use format

Poly(N-isopropylacrylamide) Hydrogels for Storage and Delivery of Reagents to Paper-Based Analytical Devices