title: Versatile carboxyl-terminus of capsid protein of porcine circovirus type 2 were recognized by monoclonal antibodies with pluripotency of binding.
abstract: We designed the peptide (C3) mimetic carboxyl-terminus (Cterminus) of capsid protein of porcine circovirus type 2b (PCV2b-1A/1B) inducing humoral immunity and generating hybridomas. The positive reactivity of the mAbs to PCV2 capsid protein was demonstrated by Western blot assay. Those mAbs also showed positive signals on PCV2b infected swine lymphocytes by indirect immunofluorescence staining. The mAb 1H3 bound to three minimal linear epitopes (P62, DPPLNP; P67, DPPLNPK; P73, LKDPPLKP), which was located at Cterminus of the capsid protein of PCV2b-1A/1B, PCV2b-1C, and PCV2a-2A respectively. The mAbs 3B2 bound to only one minimal linear epitopes (P59, KDPPLNP). The mAbs 6B8 bound to two minimal linear epitopes (P59 and P67). Our data demonstrate the core motif (P62) within the P59 could be recognized by mAbs (3B2 and 6B8) in the free status by liquid phase blocking immunoassay (LPBI) but not be recognized by these mAbs in the fixed form on the plate by indirect ELISA (iELISA). However, the P73 could be recognized by mAb 1H3 by iELISA but no inhibition of the interactive binding of C3 and mAb 1H3 by LPBI. This study also indicated that IgM mAbs and defective Ig mAb have broad binding, moderate specificity and low affinity. This study confirm that mAbs have pluripotency of binding. It might be a phenomenon of antibody response to Cterminus of capsid protein of PCV2b.
authors.affiliation: Animal Health Research Institute, Council of Agriculture, Executive Yuan, New Taipei, Taiwan; Livestock Research Institute, Council of Agriculture, Executive Yuan, Tainan, Taiwan; School of Veterinary Medicine, National Taiwan University, Taipei, Taiwan. Electronic address: lchung@mail.nvri.gov.tw.
authors.forename: Ling-Chu
authors.initials: LC
authors.lastname: Hung
authors.affiliation: School of Veterinary Medicine, National Taiwan University, Taipei, Taiwan. Electronic address: ivancheng@ntu.edu.tw.
authors.forename: Ivan-Chen
authors.initials: IC
authors.lastname: Cheng
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology*; D001666:Binding Sites, Antibody / Q000276:immunology; D015153:Blotting, Western; D036022:Capsid Proteins / Q000276:immunology*; D017925:Circovirus / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000276:immunology; D005455:Fluorescent Antibody Technique; D007118:Immunoassay; D007074:Immunoglobulin G / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D013552:Swine
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D018985:Epitopes, B-Lymphocyte; D007074:Immunoglobulin G; D007075:Immunoglobulin M
keywords: Carboxyl-terminus of the capsid protein; Fine epitope; Monoclonal antibodies; Pluripotency of binding; Porcine circovirus type 2