Identification of a Novel Conserved B Cell Epitope in the N Protein of Equine Arteritis Virus (Bucyrus Strain).
The nucleocapsid (N) protein is the most conserved structural protein in equine arteritis virus (EAV). This study aimed to identify the minimal conserved B cell epitope on the EAV N protein. The purified N protein was used to immunize mice for preparing monoclonal antibody (mAb). The reactivity of mAb was evaluated by Western blot and immunofluorescence assay. Moreover, 11 overlapping peptides (named MBP-N1 to MBP-N11) were designed to localize the linear antigenic epitope within the N protein. The peptides were identified by indirect enzyme-linked immunosorbent assay (ELISA) and Western blot. The minimal conserved B cell epitope on the EAV N protein was identified. The homology analysis was also performed. An EAV N-reactive mAb was selected and designated as 1C11. Indirect ELISA results showed that overlapping domain between MBP-N10 and MBP-N11 was recognized by the mAb 1C11. Furthermore, the indirect ELISA and Western blot showed that (101)QRKVAP(106) was the minimal linear epitope of the EAV N protein. The homology analysis showed that the identified epitope was conserved among all EAV strains analyzed in this work, with the exception of the ARVAC. One EAV N-specific mAb (1C11) was developed, and a minimal linear peptide epitope ((101)QRKVAP(106)) within the N protein was identified.
1 Department of Gynecology, The Third Affiliated Hospital of Harbin Medical University , Harbin, China .
Jie
J
Chen
2 Department of General Surgery, The Fourth Affiliated Hospital of Harbin Medical University , Harbin, China .
Xinggang
X
Guo
1 Department of Gynecology, The Third Affiliated Hospital of Harbin Medical University , Harbin, China .
Lianwei
L
Li
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000276:immunology / Q000302:isolation & purification; D015153:Blotting, Western; D017124:Conserved Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D007240:Equartevirus / Q000235:genetics / Q000276:immunology*; D005260:Female; D005455:Fluorescent Antibody Technique; D008807:Mice, Inbred BALB C; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000276:immunology*; D017386:Sequence Homology, Amino Acid
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D018985:Epitopes, B-Lymphocyte; D019590:Nucleocapsid Proteins