Functional and topographical analyses of epitopes on bovine herpesvirus type 1 glycoprotein IV.
Bovine herpesvirus type 1 (BHV-1) glycoprotein gIV was purified by affinity chromatography. Purified preparations showed two distinct components of 71 K and 140 K following electrophoresis in sodium dodecyl sulphate polyacrylamide gels. The polypeptides were separated, excised from the gel and used to immunize rabbits; the resulting antisera showed a high degree of cross reactivity indicating that these polypeptides represent monomeric and dimeric forms of the same glycoprotein. Purified gIV was also used to develop a gIV-specific panel of monoclonal antibodies. Neutralizing monoclonal antibodies directed against gIV were conjugated to horseradish peroxidase and subjected to competition binding assays by ELISA. Three distinct neutralizing antigenic domains on gIV were identified. Domain 1 comprised two overlapping epitopes, whereas domain 2 was represented by a single monoclonal antibody. The third antigenic domain was made up of a complex of four identical or overlapping epitopes designated 3a, b, c, and d. Evidence is presented suggesting that domain 1 of gIV may be involved in penetration of the virus into the cell.
Hughes
G
G
Veterinary Infectious Disease Organization, Saskatoon, Saskatchewan, Canada.
Babiuk
L A
LA
van Drunen Littel-van den Hurk
S
S
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000302:isolation & purification; D001667:Binding, Competitive; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000302:isolation & purification; D007242:Herpesvirus 1, Bovine / Q000472:pathogenicity / Q000648:ultrastructure*; D008970:Molecular Weight; D009500:Neutralization Tests; D014764:Viral Proteins / Q000302:isolation & purification / Q000502:physiology* / Q000648:ultrastructure; D014776:Virus Cultivation
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D000939:Epitopes; D014764:Viral Proteins; C046998:bovine herpesvirus type-1 glycoproteins