Versatile and efficient fabrication of signal “turn‐on” lateral flow assay for ultrasensitive naked eye detection of small molecules based on self‐assembled fluorescent gold nanoclusters‐antigen aggregates

Aggregate Pub Date : 2024-08-22 DOI:10.1002/agt2.644
Mengjia Chao, Shengmei Tai, Minxin Mao, Wenbo Cao, Chifang Peng, Wei Ma, Yongwei Feng, Zhouping Wang
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Abstract

Fluorescence signal “turn‐on” lateral flow immunoassay (FONLFA) through nanomaterial labeled quenching fluorescent nanomaterial has shown significant potential for the detection of small molecules. However, the fluorescent nanomaterial immobilization on nitrocellulose (NC) membrane commonly requires tedious chemical modification and only a few combinations of fluorescence donor and quencher have been applied in FONLFA. In this work, bright fluorescent metal nanoclusters (Prot‐AuNCs) were prepared and self‐assembled into Prot‐AuNCs/antigen aggregates with three typical small molecule antigens, respectively. The aggregates can be readily immobilized on the surface of the NC membrane, indicating that this strip fabrication strategy has good versatility. Moreover, we evaluated the performances of this FONLFA platform by using carbendazim as a model target and investigated four typical nanomaterials as colorimetric nanoprobes and fluorescence quenchers. We found that all the nanoprobes demonstrated significantly improved naked eye detection sensitivity (vLOD) and limits of detection (LODs) in quantitative analysis. Among them, combing the Fe‐polydopamine nanoparticles as quencher with the above aggregates, the FONLFA in signal “turn‐on” mode achieved 200‐fold improved vLOD (0.05 ng mL−1) compared with conventional colorimetric AuNPs‐based lateral flow immunoassay (AuNPs‐LFA) (10 ng mL−1). In addition, the LOD in quantitative analysis also was improved by 22‐fold and the whole test process was completed within 10 min. With the advantages of efficient fabrication, extraordinary sensitization, and good biocompatibility, our FONLFA platform is expected to have great potential in the rapid detection of various small molecules.

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基于自组装荧光金纳米簇-抗原聚集体的多功能、高效的信号 "开启 "横向流动分析仪,用于超灵敏肉眼检测小分子
通过纳米材料标记淬灭荧光纳米材料的荧光信号 "开启 "横向流动免疫分析法(FONLFA)已在小分子检测方面显示出巨大的潜力。然而,将荧光纳米材料固定在硝酸纤维素(NC)膜上通常需要进行繁琐的化学修饰,而且只有少数荧光供体和淬灭剂的组合被应用于 FONLFA。本研究制备了明亮的荧光金属纳米团簇(Prot-AuNCs),并分别与三种典型的小分子抗原自组装成Prot-AuNCs/抗原聚集体。这些聚集体可以很容易地固定在 NC 膜表面,表明这种条带制造策略具有良好的通用性。此外,我们还以多菌灵为模型靶标评估了该 FONLFA 平台的性能,并研究了四种典型的纳米材料作为比色纳米探针和荧光淬灭剂。我们发现,所有纳米探针在定量分析中的裸眼检测灵敏度(vLOD)和检测限(LOD)都有明显提高。其中,将多巴胺铁纳米粒子作为淬灭剂与上述聚集体相结合,在信号 "开启 "模式下,FONLFA 的 vLOD(0.05 ng mL-1)比传统的比色法 AuNPs 侧向流动免疫分析法(AuNPs-LFA)(10 ng mL-1)提高了 200 倍。此外,定量分析的 LOD 也提高了 22 倍,整个测试过程在 10 分钟内完成。我们的 FONLFA 平台具有高效制备、超强敏化和良好生物相容性等优点,有望在快速检测各种小分子化合物方面发挥巨大潜力。
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