Development of a specific nanobody and its application in rapid and selective determination of Salmonella Enteritidis in milk

Abstract The variable domain of heavy chain only antibody (VHH), also named as nanobody obtained from camelid antibody libraries, has shown great potential in immunochemistry, with many advantages over conventional antibodies. Here, nanobodies towards Salmonella Enteritidis (S. Enteritidis) were isolated for the first time after biopanning of an immune camelid nanobody library. Those nanobodies showed high thermostability and good specificity. A sandwich-ELISA was developed with nanobody Nb 13, which exhibited a detection limit of 1.4×105 CFU/mL. The proposed assay was also applied to detect S. Enteritidis in milk samples, which could detect 6 CFU/mL of S. Enteritidis in milk after 10 h of enrichment. This study demonstrated an alternative strategy for S. Enteritidis determination, and further proved the utility and practicality of nanobody-based reagents in bioanalytical chemistry.