Wang Tao, Ran Chuanjiang, He Xinyue, Li Shengzhou, Xiang Hongguang, Shen Yan, Wang Jue, Wei Hongxia
Department of Clinical Laboratory, Second People's Hospital of Taixing City Jiangsu Province 225400 China.
Department of Pharmaceutics, School of Pharmacy, China Pharmaceutical University Nanjing 210019 Jiangsu Province China.
RSC Adv. 2024 Sep 23;14(41):30154-30164. doi: 10.1039/d4ra05277c. eCollection 2024 Sep 18.
In recent years, with the rapid development of point-of-care testing, the application of lateral flow immunochromatography assay (LFIA) has become increasingly widespread. The key to the success of these detection technologies is the effective binding with diagnostic materials and detection antibody proteins. Although many researchers have tried to optimize antibody binding, a universally accepted strategy that can provide maximum performance has not been determined. In this study, the HIV infection P24 antigen was selected as the detection biomarker. Then the binding mechanism between hollow gold nanoparticles as diagnostic materials and detection antibodies was explored through dynamic light scattering, Fourier transform infrared spectroscopy, circular dichroism spectroscopy, and other methods. It was found that the binding efficiency is related to the change in protein secondary conformation during binding, hydrogen bonding, and van der Waals force maintain the binding mechanism between antibodies and nanoparticles. The main forces of particle complexation and the main binding site of the antibody were discussed and analyzed. Finally, an immunochromatographic system was constructed to evaluate the significant advantages of this platform compared to the common colloidal gold immunochromatographic system.
近年来，随着即时检测的快速发展，侧向流动免疫层析分析（LFIA）的应用越来越广泛。这些检测技术成功的关键在于与诊断材料和检测抗体蛋白的有效结合。尽管许多研究人员试图优化抗体结合，但尚未确定一种能提供最佳性能的普遍接受的策略。在本研究中，选择HIV感染P24抗原作为检测生物标志物。然后通过动态光散射、傅里叶变换红外光谱、圆二色光谱等方法，探索了作为诊断材料的中空金纳米颗粒与检测抗体之间的结合机制。研究发现，结合效率与结合过程中蛋白质二级构象的变化有关，氢键和范德华力维持了抗体与纳米颗粒之间的结合机制。讨论并分析了颗粒复合的主要作用力和抗体的主要结合位点。最后，构建了免疫层析系统，以评估该平台相对于普通胶体金免疫层析系统的显著优势。
