Mapping of linear antibody epitopes of the glycoprotein of VHSV, a salmonid rhabdovirus.
Antibody linear epitopes of the glycoprotein G (gpG) of the viral haemorrhagic septicaemia virus (VHSV), a rhabdovirus of salmonids, were mapped by pepscan using overlapping 15-mer peptides covering the entire gpG sequence and ELISA with polyclonal and monoclonal murine and polyclonal trout antibodies. Among the regions recognized in the pepscan by the polyclonal antibodies (PAbs) were the previously identified phosphatidylserine binding heptad-repeats (Estepa & Coll 1996; Virology 216:60-70) and leucocyte stimulating peptides (Lorenzo et al. 1995; Virology 212:348-355). Among 17 monoclonal antibodies (MAbs), only 2 non-neutralizing MAbs, 110 (aa 139-153) and IP1H3 (aa 399-413), could be mapped to specific peptides in the pepscan of the gpG. Mapping of these MAbs was confirmed by immunoblotting with recombinant proteins and/or other synthetic peptides covering those sequences. None of the neutralizing MAbs tested reacted with any of the gpG peptides. Previously mapped MAb resistant mutants in the gpG did not coincide with any of the linear epitopes defined by the pepscan strategy, suggesting the complementarity of the 2 methods for the identification of antibody recognition sites.
Fernandez-Alonso
M
M
INIA, Sanidad Animal, CISA-Valdeolmos, Madrid, Spain.
Lorenzo
G
G
Perez
L
L
Bullido
R
R
Estepa
A
A
Lorenzen
N
N
Coll
J M
JM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000737:chemistry / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D018604:Epitope Mapping; D000939:Epitopes / Q000032:analysis*; D005260:Female; D005393:Fish Diseases / Q000821:virology; D005434:Flow Cytometry / Q000662:veterinary; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology*; D007106:Immune Sera / Q000276:immunology; D015151:Immunoblotting / Q000662:veterinary; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests / Q000662:veterinary; D012209:Rhabdoviridae / Q000276:immunology*; D018353:Rhabdoviridae Infections / Q000662:veterinary / Q000821:virology; D012487:Salmonidae / Q000821:virology*; D014764:Viral Proteins / Q000737:chemistry / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D006023:Glycoproteins; D007106:Immune Sera; D014764:Viral Proteins