title: Identification of three novel linear B-cell epitopes on the VP5 protein of BTV16.
abstract: Bluetongue virus (BTV) VP5 protein is an important antigenic protein which is centrally involved in serotype determination and the virus entry process. Very little is known about the B-cell epitopes on the BTV VP5 protein recognized by humoral immune responses. In this study, we generated five BTV16 VP5 protein-specific monoclonal antibodies (MAbs), named 3B11, 2B10, 1H7, 4A6 and 3G9, and defined the linear epitopes recognized by MAbs using a series of peptides expressed as maltose-binding protein (MBP)-fusion polypeptides. Three novel linear B-cell epitopes were identified: 3B11 and 3G9 recognized the motif ITANTREIQHIKEE; 2B10 recognized the motif LSGID; and 4A6 recognized the motif STMVKEYRQKIDALKA. Exact sequences corresponding to the three motifs identified were found in the BTV16 VP5 protein ((310)ITANTREIQHIKEE(323), (265)LSGID(269) and (188)STMVKEYRQKIDALKA(203)). These motifs represent the minimal linear peptide sequence required for MAb reactivity, as binding of each MAb was abolished when additional amino acids were removed from the amino and carboxy termini of the peptide. Amino acid sequence alignment indicated that three epitopes were totally conserved among different BTV16 strains. The MAbs generated along with identified epitopes will be useful for examining VP5 protein function and the development of epitope-based marker vaccines against BTV.
authors.affiliation: The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China. Electronic address: srwangws@163.com.
authors.forename: Wen-Shi
authors.initials: WS
authors.lastname: Wang
authors.affiliation: The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China. Electronic address: sunlhw@gmail.com.
authors.forename: En-Cheng
authors.initials: EC
authors.lastname: Sun
authors.affiliation: The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China. Electronic address: sunencheng@yahoo.cn.
authors.forename: Ni-Hong
authors.initials: NH
authors.lastname: Liu
authors.affiliation: The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China. Electronic address: yangtao04@126.com.
authors.forename: Tao
authors.initials: T
authors.lastname: Yang
authors.affiliation: The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China. Electronic address: x_qingyuan@163.com.
authors.forename: Qing-Yuan
authors.initials: QY
authors.lastname: Xu
authors.affiliation: The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China. Electronic address: yongliqin119@163.com.
authors.forename: Yong-Li
authors.initials: YL
authors.lastname: Qin
authors.affiliation: The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China. Electronic address: zhaojingnh2005@163.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.affiliation: The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China. Electronic address: fyf_anna@163.com.
authors.forename: Yu-Fei
authors.initials: YF
authors.lastname: Feng
authors.affiliation: The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China. Electronic address: lijunping916@163.com.
authors.forename: Jun-Ping
authors.initials: JP
authors.lastname: Li
authors.affiliation: The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China. Electronic address: weipengr@sina.com.
authors.forename: Peng
authors.initials: P
authors.lastname: Wei
authors.affiliation: The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China. Electronic address: 337559885@qq.com.
authors.forename: Cui-Yun
authors.initials: CY
authors.lastname: Zhang
authors.affiliation: The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China. Electronic address: dlwu@hvri.ac.cn.
authors.forename: Dong-Lai
authors.initials: DL
authors.lastname: Wu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000918:Antibody Specificity; D036022:Capsid Proteins / Q000276:immunology*; D045744:Cell Line, Tumor; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000032:analysis / Q000276:immunology*; D005260:Female; D006168:Guinea Pigs; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009101:Multiple Myeloma / Q000821:virology; D010446:Peptide Fragments / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D017386:Sequence Homology, Amino Acid
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036022:Capsid Proteins; D018985:Epitopes, B-Lymphocyte; D010446:Peptide Fragments; D011994:Recombinant Proteins; C442520:VP2 protein, Bluetongue virus; C488468:VP5 protein, Bluetongue virus