Screening of a small set of random peptides: a new strategy to identify synthetic peptides that mimic epitopes.
Small diversity libraries, composed of 4550 synthetic dodecapeptides and 8000 synthetic tripeptides, have been used to identify sequences homologous to small linear and non-linear parts of epitopes. Here we report that synthetic peptides identified through alignment of dodecapeptides and tripeptides derived from these small libraries have, in direct ELISA and/or competitive ELISA, activities similar to that of peptides covering the native epitope and similar to that of peptides derived from large expression libraries composed of 10(6)-10(7) random peptides. This result was obtained with the monoclonal antibodies 6A.A6 and M2. Mab 6A.A6 binds the transmissible gastroenteritis virus (TGEV) and mAb M2 binds the FLAG-peptide, an affinity tag. It was also found that the antibody binding activity of peptides, derived from small or large libraries, can strongly depend on the way in which the peptide is presented to the antibody, i.e. high antibody titers were obtained when these peptides were synthesized on pins or coated onto microtiter plates, whereas low IC50s were obtained with these peptides in solution. We postulate that small peptide libraries may be a powerful tool to quickly identify new peptides that can be used as sensitive markers for mAbs of interest.
Slootstra
J W
JW
Department of Molecular Recognition, Institute for Animal Science and Health (ID-DLO), Lelystad, The Netherlands.
Puijk
W C
WC
Ligtvoet
G J
GJ
Kuperus
D
D
Schaaper
W M
WM
Meloen
R H
RH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis* / Q000276:immunology; D018716:Molecular Mimicry; D009842:Oligopeptides; D010446:Peptide Fragments / Q000032:analysis* / Q000138:chemical synthesis / Q000276:immunology*; D010455:Peptides / Q000276:immunology; D005760:Transmissible gastroenteritis virus / Q000276:immunology; D014764:Viral Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D009842:Oligopeptides; D010446:Peptide Fragments; D010455:Peptides; D014764:Viral Proteins; C064484:spike protein S, Transmissible gastroenteritis virus; C085706:FLAG peptide