Rodriguez Barroso Laura G, Lanzagorta Garcia Eduardo, Mojicevic Marija, Alkan Tas Buket, Huerta Miriam, Pogue Robert, Devine Declan M, Brennan-Fournet Margaret
PRISM Research Institute, Technological University of the Shannon: Midlands Midwest, Dublin Rd, N37 HD68 Athlone, Ireland.
Physics Institute, Universidad Autónoma de San Luis Potosí, Av. Parque Chapultepec 1570, San Luis Potosí 78295, Mexico.
Int J Mol Sci. 2023 Jul 26;24(15):11974. doi: 10.3390/ijms241511974.
Nanotechnology offers new possibilities in molecular diagnostics, with nanoparticles gaining attention as biosensor upgrades. This study evaluates gold-coated silver nanoplates coated with PEG for enhanced protection, aiming to detect Spike protein with higher sensitivity, and emphasizes the importance of considering complex environments and appropriate controls for specific binding and accurate analysis. The sensitivity of antibody-coated PEGAuTSNPs as tools for immunoassays is demonstrated through fibronectin (Fn)- anti-fibronectin binding within an isolated extracellular matrix as a complex and native environment of Fn. Moreover, the optimal functionalization volume of Spike protein was determined (4 µg/mL of PEGAuTSNP). Anti-Spike was added to confirm binding, while the TJP1 protein was used as a negative control. The same experiment was used in the presence of horse serum to simulate a complex environment. According to Localized Surface Plasmon Resonance analysis and Dynamic Light Scattering size measurements, anti-Spike exhibited a stronger affinity for the nanoplates, causing TJP1 to be replaced by the antibody on the nanoplates' surface. Future research will involve exploring alternative complex environments, filtering larger molecules, and the optimization of immunoassay performance.
纳米技术为分子诊断带来了新的可能性,纳米颗粒作为生物传感器的升级版受到关注。本研究评估了涂有聚乙二醇(PEG)以增强保护作用的金包银纳米片,旨在以更高的灵敏度检测刺突蛋白,并强调了在特定结合和准确分析中考虑复杂环境和适当对照的重要性。通过纤连蛋白(Fn)-抗纤连蛋白在分离的细胞外基质中作为Fn的复杂天然环境的结合,证明了抗体包被的PEGAuTSNP作为免疫测定工具的灵敏度。此外,还确定了刺突蛋白的最佳功能化体积(4 µg/mL的PEGAuTSNP)。加入抗刺突蛋白以确认结合,同时将紧密连接蛋白1(TJP1)蛋白用作阴性对照。在存在马血清的情况下进行相同实验以模拟复杂环境。根据局部表面等离子体共振分析和动态光散射尺寸测量,抗刺突蛋白对纳米片表现出更强的亲和力,导致TJP1被纳米片表面的抗体取代。未来的研究将包括探索替代的复杂环境、过滤较大分子以及优化免疫测定性能。