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Rapid and highly sensitive immunoassay using an ultra-thin immuno-wall microfluidic device with a sequential fluorescence signal increment method

We present a rapid and highly sensitive immunoassay platform based on an ultra-thin immuno-wall microfluidic device with an easy-to-perform sequential fluorescence signal increment method. The ultra-thin immuno-wall was fabricated using a special type of water-soluble photopolymer mixed with streptavidin via photolithography. During photolithography, the photopolymer formed a three-dimensional cross-linked structure, and streptavidin was immobilized in the cross-linked structure based on the click chemistry reaction. The immobilized streptavidin was used to immobilize biotin-conjugated antibodies on the cross-linked structure to capture biomarkers, forming immune complexes on the surface, known as an "immuno-wall." A sequential fluorescence signal increment method utilizes two different fluorescence-labeled antibodies with high affinity that were incubated several cycles in the immuno-wall to enhance the fluorescence signal. Moreover, an ultra-thin …

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