Preparation and identification of a monoclonal antibody against Mycoplasma hyorhinis
TIAN Rui-yu1, 4, XIONG Qi-yan1, NI Bo1, WANG Jia1, 2, WEI Yan-na1, FENG Zhi-xin1, SHAO Guo-qing1, 3
1.Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences/Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture/NationalCenter for Engineering Research of Veterinary Bio-products, Nanjing 210014, China; 2.Key Laboratory of Food Quality and Safety of Jiangsu Province-State Key Laboratory Breeding Base,Nanjing 210014, China; 3.Jiangsu Collaborative Innovation Center for Meat Production, Processing and Quality Control,Nanjing 210095, China; 4.College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
Abstract：The aim of this study was to produce a monoclonal antibody against Mycoplasma hyorhinis, which would be useful in the diagnosis and pathogenesis researches of M. hyorhinis. BALB/c mice were immunized with whole cell protein of M. hyorhinis, and then the monoclonal antibody (McAb) was prepared by hybridoma technique. The isotype of the McAb was identified, and then the titer and specificity were analyzed. The prepared McAb was used to detect M. hyorhinis in the colony immunoboltting assay and indirect immunofluorescence assay. A hybridoma cell line was obtained, and was named Mhr-08. The McAb belonged to IgG1 isotype, and the light chain was κ type. The titer was detected to be 1∶102 400 by ELISA. The result of Western-blot indicated that this McAb strongly reacted to a 43 kDa protein of M. hyorhinis, without cross-reaction with other swine mycoplasmas, Escherichia coli or KM2 medium. It revealed that the McAb recognized a surface membrane protein and was successfully applied to detect M. hyorhinis in colony immunoboltting assay. The mycoplasmas bound to the tracheal epithelial cells was successfully detected by using this McAb in indirect immunofluorescence assay. In conclusion, a McAb against M. hyorhinis was successfully prepared, which provides as a useful tool for the diagnosis and pathogenesis researches.
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2017, Vol. 33 
