Li Xiang, Yu Dong, Li Huiwen, Sun Ruichang, Zhang Zhuoran, Zhao Tianyu, Guo Gengchen, Zeng Jingbin, Wen Cong-Ying
College of Chemistry and Chemical Engineering, China University of Petroleum (East China), Qingdao, 266580, PR China.
Huangdao Customs of the People's Republic of China, 266580, PR China.
Biosens Bioelectron. 2023 Dec 1;241:115688. doi: 10.1016/j.bios.2023.115688. Epub 2023 Sep 10.
Traditional lateral flow immunoassays (LFIA) suffer from insufficient sensitivity, difficulty for quantitation, and susceptibility to complex substrates, limiting their practical application. Herein, we developed a polyethylenimine (PEI)-mediated approach for assembling high-density Au nanoshells onto FeO nanoclusters (MagAu) as LFIA labels for integrated enrichment and photothermal/colorimetric dual-mode detection of SARS-CoV-2 nucleocapsid protein (N protein). PEI layer served not only as "binders" to FeO nanoclusters and Au nanoshells, but also "barriers" to ambient environment. Thus, MagAu not only combined magnetic and photothermal properties, but also showed good stability. With MagAu, N protein was first separated and enriched from complex samples, and then loaded to the strip for detection. By observation of the color stripes, qualitative detection was performed with naked eye, and by measuring the temperature change under laser irradiation, quantification was attained free of sophisticated instruments. The introduction of FeO nanoclusters facilitated target purification and enrichment before LFIA, which greatly improved the anti-interference ability and increased the detection sensitivity by 2 orders compared with those without enrichment. Moreover, the high loading density of Au nanoshells on one FeO nanocluster enhanced the photothermal signal of the nanoprobe significantly, which could further increase the detection sensitivity. The photothermal detection limit reached 43.64 pg/mL which was 1000 times lower than colloidal gold strips. Moreover, this method was successfully applied to real samples, showing great application potential in practice. We envision that this LFIA could serve not only for SARS-CoV-2 detection but also as a general test platform for other biotargets in clinical samples.
传统的侧向流动免疫分析(LFIA)存在灵敏度不足、定量困难以及易受复杂基质影响等问题,限制了其实际应用。在此,我们开发了一种聚乙烯亚胺(PEI)介导的方法,用于将高密度金纳米壳组装到FeO纳米簇(磁金,MagAu)上,作为LFIA标记物,用于对严重急性呼吸综合征冠状病毒2(SARS-CoV-2)核衣壳蛋白(N蛋白)进行集成富集和光热/比色双模式检测。PEI层不仅作为FeO纳米簇和金纳米壳的“粘合剂”,还作为与周围环境的“屏障”。因此,磁金不仅兼具磁性和光热特性,还表现出良好的稳定性。利用磁金,首先从复杂样品中分离并富集N蛋白,然后加载到试纸条上进行检测。通过观察颜色条带,可进行肉眼定性检测;通过测量激光照射下的温度变化,无需复杂仪器即可实现定量检测。FeO纳米簇的引入促进了LFIA之前的目标纯化和富集,与未富集的情况相比,大大提高了抗干扰能力,并将检测灵敏度提高了2个数量级。此外,一个FeO纳米簇上金纳米壳的高负载密度显著增强了纳米探针的光热信号,这可以进一步提高检测灵敏度。光热检测限达到43.64 pg/mL,比胶体金试纸条低1000倍。此外,该方法成功应用于实际样品,在实践中显示出巨大的应用潜力。我们设想,这种LFIA不仅可用于SARS-CoV-2检测,还可作为临床样品中其他生物靶标的通用检测平台。
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