Peptide mimics of hapten DNP: the effect of affinity of anti-DNP monoclonal antibodies for the selection of phage-displayed mimotopes.
Biopanning of two linear (6- and 15-mer) and two constrained (10- and 17-mer) phage-displayed peptide libraries with two anti-DNP monoclonal antibodies (mAbs) selected seven unique peptide sequences using only the low affinity anti-DNP monoclonal antibody. The selected peptides contained two of 6, one of 10, two of 15 and two of 17 amino acids in length. They were all rich in hydrophobic residues. Both 15-mer peptides had antigenic regions of eight amino acids as revealed by a spot scan assay. Two of the 17-mer and one of the 10-mer peptides displayed on phage competed with free DNP for the low affinity anti-DNP mAb. These findings highlight (i) the selective power of phage displayed peptide libraries to identify peptides that mimic the shape of a small hapten molecule such as DNP, (ii) the possible preferential bias of phage libraries towards low affinity antibodies, (iii) the importance of using a panel of phage libraries for selecting peptide mimics.
Department of Pathology and Infectious Diseases, Royal Veterinary College, University of London, Royal College Street, London NW1 0TU, UK. Atakal61@hotmail.com
Atila T
AT
Kalaycioglu
Peter H
PH
Russell
Colin R
CR
Howard
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004140:Dinitrophenols / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006241:Haptens / Q000276:immunology*; D051379:Mice; D018716:Molecular Mimicry / Q000276:immunology*; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000235:genetics / Q000276:immunology; D011817:Rabbits
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D004140:Dinitrophenols; D000939:Epitopes; D006241:Haptens; D019151:Peptide Library; D010455:Peptides