A high signal amplification immunoassay method based on Fluorescein Diacetate (FDA) organic nanoparticle was proposed here. FDA nanoparticles were prepared in a ball mill and centrifugation was applied to get the size-uniformed SDS stabilized particle (FDA@SDS). Further SEM images confirmed the particle size was well dispersed in solution and particle size analysis results showed that the particle size 90% was distributed in the range of 200 nm to 500 nm. Mass Spectrometry results showed that the antibody can directly attach to the FDA@SDS particles and standard sandwich immunoassay confirmed the bioactivity of the antibody on the FDA@SDS particle surface (FDA@SDS-IgG). And then Zeta potential test results showed that the blocking reagents BSA and Casein can successfully block the unsatisfied sites on the FDA@SDS-IgG surface, which resulted in the improvement of nonspecific binding and stabilization. Further experiments were carried out using the standard immunoassays which showed the analyte signal can be greatly amplified when Casein was used as block buffer in the immunoassays. The limitation of detection was lower than 10 pg/mL, which is 100 times lower than Cy3 or FITC directly labeled antibody. The improved procedure simplifies previously reported protocols and provides a simple high signal amplification immunoassay.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.