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“潜在手术刀”:一种从从头设计到得出结果的生物辅助超快速染色侧向流动免疫分析方法

"Potential Scalpel": A Bioassisted Ultrafast Staining Lateral Flow Immunoassay from De Novo to Results.

作者信息

Jia Conghui, Shi Longhua, Li Yuechun, Tian Yanli, Liu Sijie, Wang Shaochi, Liao Xingrui, Wu Haofen, Wang Ziqi, Sun Jing, Zhang Daohong, Zhu Mingqiang, Ni Yongsheng, Wang Jianlong

机构信息

College of Food Science and Engineering, Northwest A&F University, 22 Xinong Road, Yangling 712100, Shaanxi, China.

Qinghai Key Laboratory of Qinghai-Tibet Plateau Biological Resources, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, 23 Xinning Road, Xining 810008, Qinghai, China.

出版信息

Anal Chem. 2023 Feb 28;95(8):4095-4103. doi: 10.1021/acs.analchem.2c04878. Epub 2023 Feb 13.

Abstract

It is of great importance to overcome potential incompatibility problems between dyestuffs and antibodies (mAbs) for extensive commercial application of a dyestuff-chemistry-based ultrafast colorimetric lateral flow immunoassay (cLFIA). Herein, inspired by traditional staining technologies, a basic dyestuff gallocyanine (GC)-assisted biogenic "potential scalpel"-based cLFIA (GC-ABPS-based cLFIA) by employing clenbuterol (CLE) as proof-of-concept was proposed to solve a high degree of incompatibility between the same potential dyestuffs and mAbs. Goat antimouse immunoglobulin (Ab) could serve as the "potential scalpel" to form the positive potential value biomolecular network self-assemblers (BNSA) with anti-CLE mAbs (Ab) by noncovalent force. The cLFIA completed the entire detection process from de novo to detection results within 30 min thanks to the easy availability and ideal marking efficiency (≤1 min, saving 0.4-10 h) of GC. Encouragingly, the proposed ultrafast GC-ABPS-based cLFIA has also exhibited high sensitivity (0.411 ng mL) and low cost (300 times) compared with other cLFIAs. Also, the feasibility of the proposed cLFIA was demonstrated by detecting CLE in beef, pork ham, and skim milk. Finally, the proposed GC-ABPS-based cLFIA has broadened the application range of dyestuffs and provided an effective reference strategy for the application of dyestuffs in food safety monitoring.

摘要

对于基于染料化学的超快速比色侧向流动免疫分析(cLFIA)的广泛商业应用而言,克服染料与抗体(单克隆抗体)之间潜在的不相容性问题至关重要。在此,受传统染色技术启发,提出了一种基于碱性染料焦性没食子蓝(GC)辅助的生物源性“潜在手术刀”的cLFIA(基于GC-ABPS的cLFIA),以克伦特罗(CLE)作为概念验证,来解决相同潜在染料与单克隆抗体之间高度的不相容性。山羊抗小鼠免疫球蛋白(Ab)可作为“潜在手术刀”,通过非共价力与抗CLE单克隆抗体(Ab)形成正电位值的生物分子网络自组装体(BNSA)。由于GC易于获得且标记效率理想(≤1分钟,节省0.4 - 10小时),该cLFIA在30分钟内完成了从从头开始到检测结果的整个检测过程。令人鼓舞的是,与其他cLFIA相比,所提出的基于超快速GC-ABPS的cLFIA还表现出高灵敏度(0.411 ng/mL)和低成本(低300倍)。此外,通过检测牛肉、火腿和脱脂牛奶中的CLE证明了所提出的cLFIA的可行性。最后,所提出的基于GC-ABPS的cLFIA拓宽了染料的应用范围,并为染料在食品安全监测中的应用提供了有效的参考策略。

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