An Automated Microfluidic Paper-Based Analytical Device for Chemiluminescence Immunoassay

This paper introduces a high-integrated microfluidic paper-based analytical device (μPAD) that has a reliable and programmable rotary valve and automated injection device. By controlling the rotation of valves, different regions on the μPAD can be connected or disconnected, allowing reagents to be effectively transported to the test zone. In order to address the limitations of traditional chemiluminescent immunoassays (CLIA), which involve expensive equipment and extensive manual operations, we utilize a smartphone to read the results and program each component of the device to automate the detection process. As a proof-of-concept, we conducted the detection of rabbit IgG under optimized experimental conditions (H2O2 concentration at 0.1 M, HRP-conjugated antibody concentration at 150 μg/mL, and plasma treatment time of 4 minutes), achieving a limit of detection of 3.58 pM. Our device combines the multifunctionality of μPAD with the sensitivity and specificity of CLIA, which potentially advances the development of point-of-care testing.