caa a22 4500 605505713 CHVBK 20210128100623.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s00253-015-6790-z doi (NATIONALLICENCE)springer-10.1007/s00253-015-6790-z Identification of three PPV1 VP2 protein-specific B cell linear epitopes using monoclonal antibodies against baculovirus-expressed recombinant VP2 protein [Elektronische Daten] [Jianhui Sun, Liping Huang, Yanwu Wei, Yiping Wang, Dongjie Chen, Wenjuan Du, Hongli Wu, Li Feng, Changming Liu] Porcine parvovirus type 1 (PPV1) is a major causative agent of embryonic and fetal death in swine. The PPV1 VP2 protein is closely associated with viral immunogenicity for eliciting neutralizing antibodies, but its antigenic structures have been largely unknown. We generated three monoclonal antibodies (MAbs) against baculovirus-expressed recombinant PPV1 VP2 protein. A PEPSCAN analysis identified the minimal B cell linear epitopes of PPV1 VP2 based on these MAbs. Three core epitopes, 228QQITDA233, 284RSLGLPPK291, and 344FEYSNGGPFLTPI356, were defined and mapped onto three-dimensional models of the PPV1 virion and VP2 monomer. The epitope 228QQITDA233 is exposed on the virion surface, and the other two are located inside the protein. An alignment of the PPV1 VP2 amino acid sequences showed that 284RSLGLPPK291 and 344FEYSNGGPFLTPI356 are absolutely conserved, whereas 228QQITDA233 has a single substitution at residue 233 in some (S→A or T). We developed a VP2 epitope-based indirect enzyme-linked immunosorbent assay (iELISA) to test for anti-PPV1 antibodies. In a comparative analysis with an immunoperoxidase monolayer assay using 135 guinea pig sera, the VP2-epitope-based iELISA had a concordance rate of 85.19%, sensitivity of 83.33%, and specificity of 85.47%. MAb 8H6 was used to monitor VP2 during the PPV1 replication cycle in vitro with an indirect immunofluorescence assay, which indicated that newly encapsulated virions are released from the nucleus at 24h postinfection and the PPV1 replication cycle takes less than 24h. This study provides valuable information clarifying the antigenic structure of PPV1 VP2 and lays the foundations for PPV1 serodiagnosis and antigen detection. Springer-Verlag Berlin Heidelberg, 2015 Porcine parvovirus type 1 nationallicence VP2 protein nationallicence MAb nationallicence Core epitope nationallicence Serodiagnosis nationallicence Sun Jianhui Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut Huang Liping Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut Wei Yanwu Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut Wang Yiping Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut Chen Dongjie Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut Du Wenjuan Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut Wu Hongli Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut Feng Li Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut Liu Changming Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/21(2015-11-01), 9025-9036 0175-7598 99:21<9025 2015 99 253 https://doi.org/10.1007/s00253-015-6790-z text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6790-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sun Jianhui Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut NATIONALLICENCE 700 1- Huang Liping Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut NATIONALLICENCE 700 1- Wei Yanwu Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut NATIONALLICENCE 700 1- Wang Yiping Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut NATIONALLICENCE 700 1- Chen Dongjie Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut NATIONALLICENCE 700 1- Du Wenjuan Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut NATIONALLICENCE 700 1- Wu Hongli Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut NATIONALLICENCE 700 1- Feng Li Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut NATIONALLICENCE 700 1- Liu Changming Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), 427 Maduan Street, Nangang District, 150001, Harbin, People's Republic of China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/21(2015-11-01), 9025-9036 0175-7598 99:21<9025 2015 99 253