Expression of Porcinized Antibody Against Cap Protein of Porcine circovirus Type 2d
WU Hao, LIU Xing-Yu, GAN Shi-Qi, ZHOU Xiao-Jie, ZHOU Ying-Shan, SONG Hou-Hui, ZHOU Xing-Dong*, WANG Xiao-Du*
College of Animal Science and Technology/College of Veterinary Medicine/Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province/Zhejiang Provincial Engineering Laboratory for Animal Health Inspection/Internet Technology, Zhejiang International Science and Technology Cooperation Base for Veterinary Medicine and Health Management/China-Australia Joint Laboratory for Animal Health Big Data Analytics, Zhejiang A&F University, Hangzhou 311300, China
AbstractPorcine circovirus type 2d (PCV2d) is a main subtype of PCV. It is an important pathogen causing multiple system failure syndrome of weaned piglets (Sus scrofa) and other pig immunosuppressation-related diseases, which seriously endangers the healthy development of China's pig industry. At present, the neutralizing epitopes of PCV2d reported are all located in the Cap protein, which is the key protein to produce neutralizing antibody. Neutralizing antibody can bind to the surface antigen of PCV2d to prevent virus infection. In this study, PCV2d Cap recombinant protein was expressed and purified by prokaryotic expression, and then immunized BALB/c mice (Mus musculus). Two hybridoma cell lines (IB3, IB4) with murine monoclonal antibodies against PCV2d Cap protein were screened, among which 1B3 had neutralizing activity and no cross-reaction with other viruses. After chimeric 1B3 antibody variable region gene and porcine constant region gene were cloned into the eukaryotic expression vector, a stable expression cell line of PCV2d Cap recombinant porcine chimeric antibody r1B3swine was prepared by Chinese hamster (Cricetulus griseus) ovarian CHO-S cells. Neutralization test and Western blot results showed that r1B3swine combined well with PCV2d virus and did not cross-react with the murine secondary antibody. In conclusion, the eukaryotic expression system of pig-derived monoclonal antibody r1B3swine against PCV2d Cap protein constructed in this study provides a material basis for the study of the structure and function of PCV2d Cap protein and the development of a new therapeutic and diagnostic formulation for PCV2.
WU Hao,LIU Xing-Yu,GAN Shi-Qi, et al. Expression of Porcinized Antibody Against Cap Protein of Porcine circovirus Type 2d[J]. 农业生物技术学报, 2023, 31(12): 2580-2588.
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