研究目的
To systematically investigate the size dependence of gold nanoparticle (AuNP) tags in surface plasmon enhanced light scattering (SP-LS) biosensing for improving sensitivity in immunoassays.
研究成果
The study confirms that larger AuNPs (50-64 nm) provide higher signal enhancement in SP-LS biosensing, as predicted by simulations, with a two-step gold enhancement protocol demonstrating feasibility for further sensitivity improvements. Optimal AuNP size and bioconjugation strategies are crucial for advancing SP-LS applications in bioanalytics.
研究不足
Aggregation issues prevented the use of larger AuNPs (>64 nm) in experiments, limiting the full exploitation of predicted signal enhancements. The gold enhancement step showed lower signal enhancement than simulated, indicating room for optimization in bioconjugation and enlargement protocols.
1：Experimental Design and Method Selection:
The study combined theoretical simulations using COMSOL Multiphysics to model SP-LS and experimental validation with AuNPs of various sizes. A sandwich immunoassay format was used with AuNP tags.
2：Sample Selection and Data Sources:
AuNPs with diameters ranging from 17 to 170 nm were synthesized and characterized. Mouse anti-goat IgG and goat anti-mouse IgG antibodies were used as model systems.
3：List of Experimental Equipment and Materials:
Equipment includes Zetasizer Nano ZS for DLS, Cary 5 UV-Vis-NIR spectrophotometer, SEM for imaging, SPR setup with flow-cell, PMT for light scattering detection, and COMSOL software for simulations. Materials include HAuCl4, sodium citrate, hydroquinone, PEG molecules, antibodies, and buffers.
4：Experimental Procedures and Operational Workflow:
AuNPs were synthesized via citrate reduction, PEGylated with mixed PEG coatings, conjugated with antibodies using EDC/NHS chemistry, and characterized by UV-Vis, DLS, and dot-blot assays. SP-LS measurements were performed on functionalized SPR sensors with varying AuNP sizes and concentrations, including a gold enhancement step for size enlargement.
5：Data Analysis Methods:
Data were analyzed using manufacturer's software for DLS, UV-Vis spectra analysis, and COMSOL simulations for electromagnetic field and scattered power calculations. Statistical analysis included size measurements and signal intensity comparisons.
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