Utilisation of bacteriophage display libraries to identify peptide sequences recognised by equine herpesvirus type 1 specific equine sera.
Three filamentous phage random peptide display libraries were used in biopanning experiments with purified IgG from the serum of a gnotobiotic foal infected with equine herpesvirus-1 (EHV-1) to enrich for epitopes binding to anti-EHV-1 antibodies. The sequences of the amino acids displayed were aligned with protein sequences of EHV-1, thereby identifying a number of potential antibody binding regions. Presumptive epitopes were identified within the proteins encoded by genes 7 (DNA helicase/primase complex protein), 11 (tegument protein), 16 (glycoprotein C), 41 (integral membrane protein), 70 (glycoprotein G), 71 (envelope glycoprotein gp300), and 74 (glycoprotein E). Two groups of sequences, which aligned with either glycoprotein C (gC) or glycoprotein E (gE), identified type-specific epitopes which could be used to distinguish between sera from horses infected with either EHV-1 or EHV-4 in an ELISA using either the phage displaying the peptide or synthetic peptides as antigen. The gC epitope had been previously identified as an immunogenic region by conventional monoclonal antibody screening whereas the gE antibody binding region had not been previously identified. This demonstrates that screening of phage display peptide libraries with post-infection polyclonal sera is a suitable method for identifying diagnostic antigens for viral infections such as EHV-1.
Animal Health Trust, Centre for Preventive Medicine, Newmarket, UK.
I
I
Birch-Machin
S
S
Ryder
L
L
Taylor
P
P
Iniguez
M
M
Marault
L
L
Ceglie
S
S
Zientara
C
C
Cruciere
F
F
Cancellotti
G
G
Koptopoulos
J
J
Mumford
M
M
Binns
N
N
Davis-Poynter
D
D
Hannant
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology*; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D001435:Bacteriophages / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D006566:Herpesviridae Infections / Q000276:immunology / Q000662:veterinary / Q000821:virology; D004861:Herpesvirus 1, Equid / Q000235:genetics / Q000276:immunology*; D006734:Horse Diseases / Q000276:immunology / Q000821:virology; D006736:Horses; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D000914:Antibodies, Viral; D000956:Antigens, Viral; D019151:Peptide Library; D010455:Peptides