Pentameric nanobodies serve as a capture agent and RANbodies function as immunoprobes for the sensitive detection of Salmonella typhimurium in immunoassays.

IF 3.8 2区 化学 Q1 BIOCHEMICAL RESEARCH METHODS
Yuan Weng, Ming Yang, Yunmin Wang, Qiang Xu, Miwan Shi, Aomo Tan, Peiling Yuan, Changwei Lei, Kui Gu
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引用次数: 0

Abstract

A simpler, more convenient, and faster diagnostic method for detection of Salmonella typhimurium (S. typhimurium) is urgently needed. In this study, the pentameric nanobodies (pST-Nb4) and HRP fusion nanobodies (ST-Nb1-vHRP) were produced in vitro, and pST-Nb4 displayed a binding affinity to S. typhimurium that was 61.8 times higher than that of the monomeric ST-Nb4. Ultimately, the sandwich ELISA based on pST-Nb4 as a capture agent and ST-Nb1-vHRP as an immunoprobe was developed for rapid detection of S. typhimurium. The assay was validated to have good specificity with a lower detection limit of 2.37 × 104 CFU/mL and a recovery rate of 95.00 to 108.66% for detection of S. typhimurium from contaminated milk and egg samples. After bacterial enrichment, the developed immunoassay was able to detect less than 10 CFU of S. typhimurium in milk or egg samples, even in mouse tissues infected with S. typhimurium.

在免疫分析中,五聚体纳米体作为捕获剂,ranbody作为免疫探针用于鼠伤寒沙门菌的敏感检测。
目前迫切需要一种更简单、更方便、更快速的鼠伤寒沙门菌诊断方法。本研究在体外制备了五聚体纳米体(pST-Nb4)和HRP融合纳米体(ST-Nb1-vHRP),发现pST-Nb4对鼠伤寒沙门氏菌的结合亲和力是单聚体ST-Nb4的61.8倍。最终,建立了以pST-Nb4为捕获剂,ST-Nb1-vHRP为免疫探针的夹心ELISA快速检测鼠伤寒沙门氏菌。结果表明,该方法特异性较好,检出限为2.37 × 104 CFU/mL,回收率为95.00 ~ 108.66%。在细菌富集后,开发的免疫分析法能够在牛奶或鸡蛋样品中检测到少于10 CFU的鼠伤寒沙门氏菌,甚至在感染鼠伤寒沙门氏菌的小鼠组织中也是如此。
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来源期刊
CiteScore
8.00
自引率
4.70%
发文量
638
审稿时长
2.1 months
期刊介绍: Analytical and Bioanalytical Chemistry’s mission is the rapid publication of excellent and high-impact research articles on fundamental and applied topics of analytical and bioanalytical measurement science. Its scope is broad, and ranges from novel measurement platforms and their characterization to multidisciplinary approaches that effectively address important scientific problems. The Editors encourage submissions presenting innovative analytical research in concept, instrumentation, methods, and/or applications, including: mass spectrometry, spectroscopy, and electroanalysis; advanced separations; analytical strategies in “-omics” and imaging, bioanalysis, and sampling; miniaturized devices, medical diagnostics, sensors; analytical characterization of nano- and biomaterials; chemometrics and advanced data analysis.
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