title: Epitope screening of the PCV2 Cap protein by use of a random peptide-displayed library and polyclonal antibody.
abstract: Capsid protein (Cap), the only structural protein of porcine circovirus 2 (PCV2), is involved in the host protective response and is a target for vaccine development. To find a rapid and easy way to fully map the antigenic epitopes of Cap, purified Cap-specific polyclonal antibodies were used to screen a random heptapeptide phage display library. After three rounds of screening, twenty phage clones that had binding activity to Cap-specific antibodies (tested by phage ELISA) were sequenced. When the inserted amino acid sequences were aligned with the Cap protein sequence, eight core regions in Cap ((50)SRTFGYT(56), (62)VRTPSW(67), (68)AVDMMR(73), (79)FLPPGG(84), (86)SNPRSVPF(93), (102)KVEFWP(107), (119)GSSXXXLDDN(128) and (229)PPLNP(233)) were identified, three of which ((50)SRTFGYT(56), (86)SNPRSVPF(93) and (102)KVEFWP(107)) for the first time. Nine phagetopes representing the eight regions were chosen to immunize Kunming mice. All except minotopes (50)SRTFGYT(56) and (229)PPLNP(233) induced antibodies against PCV2 when injected into Kunming mice.
authors.affiliation: College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan Province 410128, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Ge
authors.forename: Ai
authors.initials: A
authors.lastname: Yan
authors.forename: Wei
authors.initials: W
authors.lastname: Luo
authors.forename: Yun-Fei
authors.initials: YF
authors.lastname: Hu
authors.forename: Run-Cheng
authors.initials: RC
authors.lastname: Li
authors.forename: Da-Liang
authors.initials: DL
authors.lastname: Jiang
authors.forename: Xing-Long
authors.initials: XL
authors.lastname: Yu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D036022:Capsid Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D018173:Circoviridae Infections / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D017925:Circovirus / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D018604:Epitope Mapping; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D013552:Swine; D013553:Swine Diseases / Q000276:immunology / Q000821:virology*
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D019151:Peptide Library
keywords: Capsid protein; Epitope mapping; Phage display; Polyclonal antibodies; Porcine circovirus 2