Probing the basis of antibody reactivity with a panel of constrained peptide libraries displayed by filamentous phage.
The structural requirements for peptide binding to an antibody may be elucidated by probing it with a variety of peptides having different constraints. To this end, we have constructed and screened a panel of peptide libraries displayed by filamentous bacteriophage. The peptides in most of the libraries have the potential for constraint by fixed Cys residues, which have been placed at different sites within a randomized amino acid sequence of varying length. When taken together, the binding data obtained from screening the panel with a given antibody allow one to determine the types of constraints that promote binding, as well as the residues that are critical for binding. We describe the construction of 11, pVIII-displayed, peptide libraries, whose sizes range from 150 million to 10 billion clones. The libraries were screened with a number of polyclonal and monoclonal antibodies against peptides, proteins and carbohydrates. Cross-reactivity with peptides was always found for antibodies produced against peptides, linear epitopes on folded proteins and, surprisingly, carbohydrates, whereas antibodies against discontinuous epitopes on proteins were found less frequently. The implications of these results are discussed in terms of the structural basis for cross-reactivity with peptides.
Bonnycastle
L L
LL
Institute of Molecular Biology and Biochemistry, Simon Fraser University Burnaby, BC, Canada.
Mehroke
J S
JS
Rashed
M
M
Gong
X
X
Scott
J K
JK
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D002241:Carbohydrates / Q000276:immunology; D016384:Consensus Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D005822:Genetic Vectors / Q000235:genetics*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology*
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D000939:Epitopes; D010446:Peptide Fragments; D011994:Recombinant Proteins