期刊
ANALYTICAL BIOCHEMISTRY
卷 493, 期 -, 页码 1-7出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2015.09.023
关键词
Enzyme-linked immunosorbent assay (ELISA); Listeria monocytogenes; Naive library; Phage display; Variable domain of heavy-chain antibodies (VHHs)
资金
- Program of State Key Laboratory of Food Science and Technology [SKLF-ZZA-201302, SKLF-ZZB-201306]
- National Natural Science Foundation of China [31271863, 31301479]
- Educational Commission of Jiangxi Province [GJJ12142, GJJ13574]
- Natural Science Foundation of Jiangxi Province [20114BAB205039]
Listeria monocytogenes (LM), one of the eight species belonging to the genus Listeria, is pathogenic for both humans and animals. In this study, two novel LM-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 detect LM in pasteurized milk, with a detection limit of 1 x 10(4) colony-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. (C) 2015 Elsevier Inc. All rights reserved.
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