Peptide synthesis on Sepharose beads

NHS-activated Pharmacia HiTrap Sepharose was modified with 1, 3-diaminopropane to give an amino-functionalized support suitable for solid-phase peptide synthesis. The amide linker p-[(R1S)-alpha-[1-(9H-fluoren-9-yl)-methoxyformamido]- 2, 4-dimethoxybenzyl]phenoxyacetic acid was incorporated and the acyl carrier protein sequence 65-74 was synthesized manually on this support by the Fmoc procedure under controlled conditions with monitoring of the coupling reactions. The performance of the support in automated multiple synthesis in open reactors, with an Abimed AMS 422 according to standard protocols, was evaluated by the synthesis of the acyl carrier protein sequence 65-74 and two other 15-mer and 18-mer peptides. The quality of the resulting crude peptides was determined by HPLC and MALDI-MS, and compared with the same sequences synthesized in parallel on the commercial peptide synthesis resin TentaGel S RAM. The modified HiTrap material was found to be particularly suited for Fmoc solid-phase peptide synthesis and should be advantageous for the utilization of immobilized peptides and peptide libraries in biological assays.