Selection of new Epitopes from monovalent displayed phage octapeptide library.
A library of 2 x 10(7) random octapeptides was constructed by use of phagemid-based monovalent phage display system. The randomly synthesized degenerated oligodeoxyribonucleotides (oligos) were fused to the truncated g III (p230-p403). Sequence analysis of 11 randomly chosen clones suggested that the degenerated inserts and its deduced amino acid (aa) sequences are randomly distributed. The library was used to select binding peptides to the monoclonal antibody (mAb) 9E10, which recognizes a continuous decapeptide epitope of denatured human c-myc protein. After four to five rounds of panning, most of the eluted clones could bind to 9E10. Sequence analysis of the selected positive clones indicated that the binding sequences could fall into two classes, one class (clone 1) shares a consensus motif, ISE x x L, with c-myc decapeptide; and the sequences of the other class are entirely different. The binding of both classes to 9E10 could be specifically inhibited by free c-myc decapeptide. The immunogenicity of the phage peptide was further investigated by construction of multivalent displayed phage peptides and immunization of animals with or without adjuvant. ELISA and competitive ELISA showed that anti-serum from both mice and rabbit immunized with either clone could bind to the original antigen, c-myc decapeptide. These results denote that in spite of the dissimilarity of the selected peptides with c-myc decapeptide, they are capable of inducing similar immune respones in vivo, thus actually mimicking the antigen epitope.
Li
Q
Q
Beijing Institute for Cancer Research, Beijing 100034.
Wang
Y
Y
Li
J
J
Wang
Y
Y
Xu
J
J
Wang
L
L
Dong
Z
Z
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D002213:Capsid / Q000235:genetics*; D036022:Capsid Proteins; D016056:Immunodominant Epitopes / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D010446:Peptide Fragments / Q000235:genetics; D019151:Peptide Library; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics
D000911:Antibodies, Monoclonal; D036022:Capsid Proteins; D016056:Immunodominant Epitopes; D009838:Oligodeoxyribonucleotides; D010446:Peptide Fragments; D019151:Peptide Library; D016271:Proto-Oncogene Proteins c-myc; D011993:Recombinant Fusion Proteins; C110214:gene III protein, bacteriophage Phi-Lf