Immobilization and detection of small molecules is one of the challenging tasks in any given sensing system as the dissociation equilibrium constant is higher. Generating a right immobilization system with small molecules is mandatory for developing the drug-discovery process and disease identification. Immobilizing smaller probes on the ELISA plate is challenging because of its less adsorption on the polystyrene (PS) substrate. This research work developed an iron nanomaterial-based linker to attach osteopontin-specific aptamer on PS substrate. Iron oxide nanoparticle was attached on PS plate through amine modification and then antibody was attached by COOH reaction. On the osteopontin-modified plate, osteosarcoma biomarker of osteopontin was identified by its specific antibody and aptamer sandwich with the detection limit of 1 nM. Further, biofouling experiments with other molecules, such as lysozyme, and complementary aptamer failed to show the ELISA adsorption signal, indicating the iron oxide nanoparticle-modified PS plate specifically recognizes osteopontin. This research work effectively identifies the lesser abundance of osteopontin and helps to diagnose the osteosarcoma-related problems.
Keywords: ELISA; aptamer; nanomaterial; osteopontin; osteosarcoma.
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