Improved performances of spot multiple peptide synthesis.

We have developed a new software for the design of peptides to be prepared by the manual Spot synthesis method. It covers most of the common protocols for epitope mapping and offers flexibility for planning the experiment. We have also quantified the coupling efficiencies of three different coupling methods for the synthesis of four model decapeptides. The DIC/HOBT procedure was superior (mean coupling yield per cycle: 87% to 91%) to methods using pentafluorophenyl esters of Fmoc amino acids. All four peptides prepared by the three different coupling protocols were reactive with their cognate monoclonal antibody. Finally, using the best coupling method, we synthesized peptides of increasing length (10 to 29 residues) bearing the model epitope AcPGK at the N terminus and showed that peptides of up to 29 residues were immunoreactive.