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Enhancing Oriented Immobilization Efficiency: A One-for-Two Organism-Bispecific Nanobody Scaffold for Highly Sensitive Detection of Foodborne Pathogens.
Wang, Yueqi; Zhang, Liang; Wang, Peng; Liao, Xingrui; Dai, Yueyan; Yu, Qingyan; Yu, Gege; Zhang, Yao; Wei, Juan; Jing, Yinnan; Wang, Jiamin; Chen, Pengyu; Guo, Bing; Wang, Jianlong; Wang, Yanru.
Afiliação
  • Wang Y; College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
  • Zhang L; College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
  • Wang P; College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
  • Liao X; College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
  • Dai Y; College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
  • Yu Q; College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
  • Yu G; College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
  • Zhang Y; College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
  • Wei J; College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
  • Jing Y; College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
  • Wang J; College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
  • Chen P; College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
  • Guo B; College of Life Sciences, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
  • Wang J; College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
  • Wang Y; College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
Anal Chem ; 95(46): 17135-17142, 2023 11 21.
Article em En | MEDLINE | ID: mdl-37941297
ABSTRACT
Nanobodies have gained widespread application in immunoassays. However, their small size presents a significant challenge in achieving effective immobilization and optimal sensitivity. Here, we present a novel "one-for-two"-oriented immobilization platform based on an organism-bispecific nanobody (O-BsNb) scaffold, enabling highly sensitive detection of two bacterial pathogens. Through genetic engineering, a bispecific nanobody (BsNb) was engineered, targeting Salmonella spp. and Vibrio parahaemolyticus. The O-BsNb scaffold allowed one nanobody to bind specifically to inactivated bacteria, forming an organism-oriented immobilization platform, while the other served as the capture antibody. Consequently, the O-BsNb bioscaffold-based ELISA (O-ELISA) for individual detection of S. enteritidis and V. parahaemolyticus was established. When compared to the sandwich ELISA utilizing passive immobilization of monovalent nanobodies, the O-ELISA exhibited a remarkable 13.4- and 13.7-fold improvement in LOD for S. enteritidis and V. parahaemolyticus, respectively, highlighting the enhanced immobilization efficacy of the O-ELISA. Furthermore, the feasibility and reproducibility of the assay in practical samples were meticulously evaluated, revealing exemplary performance in terms of recovery precision and assay stability. These findings demonstrate the significant potential of the O-ELISA platform for the sensitive detection of macromolecules, opening new avenues for efficient pathogen identification in foodborne safety and clinical diagnostics.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Anticorpos de Domínio Único Idioma: En Ano de publicação: 2023 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Anticorpos de Domínio Único Idioma: En Ano de publicação: 2023 Tipo de documento: Article