Abstract

Magnetic nanoparticles (MNPs) have emerged as a promising material in disease diagnostics due to their potential to enhance detection sensitivity, facilitate concentration and purification of target substances in diverse samples, and enable favorable color-based detection. In this study, antibody-conjugated MNPs were successfully synthesized and validated through two appropriate methods: the measurement of MNPs' size and the use of phosphatase methods. Additionally, three methods were suggested and implemented for developing color in MNPs-based immunoassay, including the formation of MNP aggregations, utilization of MNPs' peroxidase-like activity, and synthesis of dually-conjugated MNPs with both enzyme and antibody. In particular, color development utilizing nanoparticle aggregations was demonstrated to result in a more yellowish color as virus concentration increased, while the peroxidase activity of MNPs exhibited a proportional increase in color intensity as the MNP concentration increased. This observation suggests the potential applicability of quantitative analysis using these methods. Furthermore, effective concentration and purification of target substances were demonstrated through the collection of MNPs using an external magnetic field, irrespective of factors such as antibody conjugation, dispersion medium, or virus binding. Finally, based on the key findings of this study, a design proposal for MNPs-based immunoassay is presented. Overall, MNPs-based immunoassays hold significant potential for advancing disease diagnostics.

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