title: Synthesis of peptide analogues using the multipin peptide synthesis method.
abstract: Modification of the multipin peptide synthesis method which allows the simultaneous synthesis of large numbers of different peptide analogues is described. Peptides were assembled on polyethylene pins derivatized with a 4-(beta-alanyloxymethyl)benzoate (beta-Ala-HMB) handle. For comparative purposes, peptides were also assembled on the diketopiperazine-forming handle N epsilon-(beta-alanyl)lysylprolyloxylactate. In model studies it was demonstrated that beta-Ala-HMB-linked peptides were cleaved from polyethylene pins with dilute sodium hydroxide or 4% methylamine/water to yield analogues with beta-Ala-free acid (beta-Ala-CO2H) and beta-Ala-methylamide (beta-Ala-CONHCH3), respectively. To assess the suitability of this approach for T-cell determinant analysis, analogues of a known T-cell determinant were synthesized with the various C-terminal endings. Peptides were characterized by amino acid analysis and fast atom bombardment-mass spectrometry. HPLC of the crude cleaved peptides indicated that 22 of the 24 peptides were greater than 95% pure. These crude peptide solutions were nontoxic in sensitive cell culture assays without further purification. All three cleavage procedures gave comparable activities in T-cell proliferation assays. These results demonstrate the potential of the multipin peptide synthesis method for the production of large numbers of different peptide analogues.
authors.affiliation: Coselco Mimotopes Pty Ltd., Clayton, Victoria, Australia.
authors.forename: R M
authors.initials: RM
authors.lastname: Valerio
authors.forename: M
authors.initials: M
authors.lastname: Benstead
authors.forename: A M
authors.initials: AM
authors.lastname: Bray
authors.forename: R A
authors.initials: RA
authors.lastname: Campbell
authors.forename: N J
authors.initials: NJ
authors.lastname: Maeji
mesh_terms: D000595:Amino Acid Sequence; D001671:Biochemistry / Q000379:methods*; D002999:Clone Cells / Q000187:drug effects; D006801:Humans; D008213:Lymphocyte Activation / Q000187:drug effects; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D011095:Polyethylenes; D013601:T-Lymphocytes / Q000187:drug effects; D015091:beta-Alanine
chemical_list: D010455:Peptides; D011095:Polyethylenes; D015091:beta-Alanine