Dot-blot immunoassay primarily based on Antibody-Nanocluster Biohybrids as Tags for naked-eye detection


Paper-based assays akin to dot-blot present excessive promise to develop point-of-care testing units fulfilling the ASSURED necessities recommended by the World Well being Group (inexpensive, delicate, particular, user-friendly, fast and sturdy, equipment-free, and deliverable). On this method, pure enzymes are conventionally employed tags to supply bioreceptors akin to antibodies with catalytic exercise for quantitative evaluation. Nonetheless, their inherent biomolecular limitations pose vital challenges, together with price and storage constraints. We suggest an alternate conjugation for antibodies primarily based on catalytic bimetallic nanoclusters, and their integration in such easy colorimetric paper-based immunoassay. The nanoclusters are composed of gold and platinum and they’re embedded within the construction of an anti-rabbit antibody, integrating in a single element the biorecognition and transduction parts required for the biosensing. The detection relies on the catalytic properties of the NCs to oxidize an insoluble chromogenic substrate, producing a visual sign on the floor of the paper that may be additional analysed for quantitative outcomes. We display the detection of antibodies towards the irritation biomarker interleukin-6 with a restrict of detection of 200 ng mL-1. Experimental outcomes reveal enhancements when it comes to stability in comparison with the pure enzyme horseradish peroxidase, retaining most of its exercise after a storage equal to six months at 4°C. Moreover, incorporating the NCs throughout the antibody construction as an alternative of attaching them by way of a covalent bond offers enhanced sensitivity of 69.7 %. This assay might be transferred to different particular antibodies to detect and quantify different analytes of curiosity.

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