Identification of a conserved linear B-cell epitope at the N-terminus of the E2 glycoprotein of Classical swine fever virus by phage-displayed random peptide library.
The E2 protein of Classical swine fever virus (CSFV) is an important envelope glycoprotein, which is responsible for inducing protective immune response in swine. Four antigenic domains, A-D, have been mapped on the E2 protein. The present study describes the identification of a linear B-cell epitope at the N-terminus of the E2 protein by screening a phage-displayed random 12-peptide library with the neutralizing monoclonal antibody (mAb) HQ06 directed against the E2 protein. HQ06 was found to bind to the phages displaying a consensus motif LFDGSNP, which is highly homologous to (772)LFDGTNP(778) of the CSFV polyprotein, locating on the border between antigenic domains B/C and A of the E2 protein. Considering that HQ06 showed reactivity with the motif (772)LFDGTNP(778) expressed as GST fusion in Western blot and indirect ELISA, we propose that the motif (772)LFDGTNP(778) represents a linear B-cell epitope of the E2 protein. The motif (773)FDGTNP(778) is the minimal requirement for the reactivity as demonstrated by analysis of the reactivity of HQ06 with several truncated peptides derived from the motif. Alignment of amino acid sequences from a number of CSFV isolates indicated that the epitope is well conserved among different subgroups of CSFV. Substitutions of the individual residues within the epitope (773)FDGTNP(778) demonstrated that residues (773)F, (775)G, and (778)P constitute the core of the epitope. The identified epitope will be useful for development of diagnostic assays and epitope-based marker vaccines against CSFV.
Peng
Wu-Ping
WP
Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, 427 Maduan Street, Harbin 150001, Heilongjiang, China.
Hou
Qiang
Q
Xia
Zhao-He
ZH
Chen
Dan
D
Li
Na
N
Sun
Yuan
Y
Qiu
Hua-Ji
HJ
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D006692:Classical Swine Fever Virus / Q000737:chemistry / Q000276:immunology*; D017124:Conserved Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry* / Q000276:immunology; D008969:Molecular Sequence Data; D019151:Peptide Library; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D018985:Epitopes, B-Lymphocyte; D019151:Peptide Library; D014759:Viral Envelope Proteins; C087086:glycoprotein E2, classical swine fever virus