Abstract |
Overlapping fragments of the bovine herpesvirus-1 (BHV-1) glycoprotein (gD) ORF were expressed as trpE-gD fusion proteins in Escherichia coli to map linear neutralizing epitopes defined by BHV-1-specific MAbs. The MAbs 3402 and R54 reacted with the expressed fragments on Western blots that located the epitopes between the amino acids 52-126 and 165-216, respectively, of gD. Bovine covalescent sera with high neutralizing antibody titers against BHV-1 reacted with these bacterially expressed proteins containing both of the epitopes. Alignment of these sequences from BHV-1 with the corresponding region of the BHV-5 gD ORF sequences (reported here) identified several amino acid mismatches. Since the MAbs 3402 and R54 neutralize the BHV-1 and not BHV-5, it was presumed that these were important amino acids in defining the epitope. To further localize the neutralizing epitopes, synthetic peptides corresponding to these regions in the BHV-1 gD ORF were tested for their capacity to block monoclonal antibody neutralization of BHV-1 infectivity. The peptides encompassing amino acids 92-106 (3402 epitope) and amino acids 202-213 (R54 epitope) of the BHV-1 gD competed with BHV-1 for the binding by MAbs 3402 and R54, respectively, in a dose-dependent manner. Antisera produced in rabbits to these peptides conjugated to a carrier reacted strongly with a 30-kDa protein by Western blotting and had neutralizing antibody titers against BHV-1.
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Authors | O Y Abdelmagid, H C Minocha, J K Collins, S I Chowdhury |
Journal | Virology
(Virology)
Vol. 206
Issue 1
Pg. 242-53
(Jan 10 1995)
ISSN: 0042-6822 [Print] United States |
PMID | 7530392
(Publication Type: Journal Article, Research Support, U.S. Gov't, Non-P.H.S., Research Support, U.S. Gov't, P.H.S.)
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Chemical References |
- Antibodies, Monoclonal
- Epitopes
- Recombinant Fusion Proteins
- Viral Envelope Proteins
- glycoprotein D, Human herpesvirus 1
- glycoprotein D, bovine herpesvirus 5
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Topics |
- Amino Acid Sequence
- Animals
- Antibodies, Monoclonal
(immunology)
- Cattle
- Cells, Cultured
- Epitopes
(chemistry, immunology)
- Herpesvirus 1, Bovine
(immunology)
- Molecular Sequence Data
- Neutralization Tests
- Open Reading Frames
- Recombinant Fusion Proteins
(immunology)
- Sequence Homology, Amino Acid
- Viral Envelope Proteins
(chemistry, immunology)
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