Peptide mimics selected from immune sera using phage display technology can replace native antigens in the diagnosis of Epstein-Barr virus infection.
There is an expanding area of small molecule discovery, especially in the area of peptide mimetics. Peptide sequences can be used to substitute for the entire native antigen for use in diagnostic assays. Our approach is to select peptides that mimic epitopes of the natural immune response to Epstein-Barr virus (EBV) that may be recognised by antibodies typically produced after infection with EBV. We screened a random peptide library on sera from rabbits immunised with a crude preparation of EBV and serum antibodies from a patient with a high titer of EBV antibodies. We selected four peptides (Eb1-4) with the highest relative binding affinity with immune rabbit sera and a single peptide with high affinity to human serum antibodies. The peptides were coupled to the carrier molecule BSA and the recognition of the peptides by IgM antibodies in clinical samples after infection with EBV was measured. The sensitivities were Eb1 94%, Eb2, 3, 4 88%, H1 81% and all had 100% specificity. This study illustrates that the phage display approach to select epitope mimics can be applied to polyclonal antibodies and peptides that represent several diagnostically important epitopes can be selected simultaneously. This panel of EBV peptides representing a wide coverage of immunodominant epitopes could replace crude antigen preparations currently used for capture in commercial diagnostic tests for EBV.
Casey
J L
JL
AdAlta Pty Ltd, 15/2 Park Drive, Bundoora, VIC 3083, Australia.
Coley
A M
AM
Parisi
K
K
Foley
M
M
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000915:Antibody Affinity; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D020031:Epstein-Barr Virus Infections / Q000175:diagnosis* / Q000276:immunology; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D018716:Molecular Mimicry / Q000235:genetics; D019151:Peptide Library; D010455:Peptides / Q000235:genetics / Q000276:immunology*; D011485:Protein Binding; D011817:Rabbits; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine
D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D019151:Peptide Library; D010455:Peptides; D012710:Serum Albumin, Bovine