Identification and characterization of species-specific nanobodies for the detection of Listeria monocytogenes in milk.
Identification and characterization of species-specific nanobodies for the detection of Listeria monocytogenes in milk.
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DOI：
10.1016/j.ab.2015.09.023
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发表时间：
2016-01
影响因子：
2.9
通讯作者：
Zhui Tu;Qi Chen;Yan-ping Li;Yonghua Xiong;Yang Xu;Na Hu;Yong Tao
中科院分区：
文献类型：
--
作者：
Zhui Tu;Qi Chen;Yan-ping Li;Yonghua Xiong;Yang Xu;Na Hu;Yong Tao
Listeria monocytogenes (LM), one of the eight species belonging to the genusListeria, is pathogenic for both humans and animals. In this study, two novelLM-specific clones, designated L5-78 and L5-79, were isolated from a phage display antibody library that was derived from the variable domain of heavy-chain antibodies (VHHs) of non-immunized alpaca. These two clones were expressed, purified, and characterized. Results showed that both isolated VHHs recognize three serotypes (1/2a, 1/2b, and 4b), which are responsible for more than 95% of documented human listeriosis cases. The recombinant VHHs possess high thermal stability, pH tolerance, and urea resistance. A sandwich enzyme-linked immunosorbent assay (ELISA) based on the VHH clone L5-79 and a monoclonal antibody was developed to detectLMin pasteurized milk, with a detection limit of 1 × 104colony-forming units (CFU)/ml. These findings indicated that the species-specific VHHs could be directly isolated from the non-immunized library with a properly designed panning strategy and VHH could be a new source for possible diagnosis/detection of foodborne pathogens in food because it was shown to be highly specific and stable.
