Yuan Weng, Ming Yang, Yunmin Wang, Qiang Xu, Miwan Shi, Aomo Tan, Peiling Yuan, Changwei Lei, Kui Gu
{"title":"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。","authors":"Yuan Weng, Ming Yang, Yunmin Wang, Qiang Xu, Miwan Shi, Aomo Tan, Peiling Yuan, Changwei Lei, Kui Gu","doi":"10.1007/s00216-025-05973-2","DOIUrl":null,"url":null,"abstract":"<p><p>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 × 10<sup>4</sup> 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.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8000,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Pentameric nanobodies serve as a capture agent and RANbodies function as immunoprobes for the sensitive detection of Salmonella typhimurium in immunoassays.\",\"authors\":\"Yuan Weng, Ming Yang, Yunmin Wang, Qiang Xu, Miwan Shi, Aomo Tan, Peiling Yuan, Changwei Lei, Kui Gu\",\"doi\":\"10.1007/s00216-025-05973-2\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>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 × 10<sup>4</sup> 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.</p>\",\"PeriodicalId\":462,\"journal\":{\"name\":\"Analytical and Bioanalytical Chemistry\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":3.8000,\"publicationDate\":\"2025-07-10\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Analytical and Bioanalytical Chemistry\",\"FirstCategoryId\":\"92\",\"ListUrlMain\":\"https://doi.org/10.1007/s00216-025-05973-2\",\"RegionNum\":2,\"RegionCategory\":\"化学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Analytical and Bioanalytical Chemistry","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1007/s00216-025-05973-2","RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
Pentameric nanobodies serve as a capture agent and RANbodies function as immunoprobes for the sensitive detection of Salmonella typhimurium in immunoassays.
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.
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.