Evaluation of peptide libraries: an iterative strategy to analyze the reactivity of peptide mixtures with antibodies.
Peptide libraries corresponding to a presumed mixture of 50,625 tetrapeptides or 16,777,216 hexapeptides were each prepared in a single assembly by standard solid-phase peptide synthesis. By enzyme-linked immunosorbent assay, the tetrapeptide library was shown to inhibit the binding of an antiserum to FMRF amide with an FLRF capture antigen; the hexapeptide library was shown to inhibit the binding of a monoclonal antibody to a 28 amino acid peptide with the corresponding peptide capture antigen. An iterative strategy of variation was used to determine for each position in the tetra- or hexapeptides which amino acid contributed the most to activity. As a result we were able to logically select out of the tetrapeptide library the sequence FLRF and to select out of the hexapeptide library a sequence that differed from the apparent probable epitope but was twice as active. A single amino acid substitution in the logically derived sequence gave a peptide that was 35 times as active as the hexapeptide sequence in the original 28 amino acid peptide.
Blake
J
J
Bristol-Myers Squibb Pharmaceutical Research Institute, Seattle, Washington 98121.
Litzi-Davis
L
L
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D004797:Enzyme-Linked Immunosorbent Assay; D019835:FMRFamide; D007124:Immunoenzyme Techniques; D007447:Invertebrate Hormones / Q000378:metabolism; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism; D010455:Peptides / Q000378:metabolism*; D011485:Protein Binding
D000911:Antibodies, Monoclonal; D007447:Invertebrate Hormones; D009479:Neuropeptides; D009842:Oligopeptides; D010455:Peptides; D019835:FMRFamide; C066148:phenylalanyl-leucyl-arginyl phenylalaninamide