Semi-automated microwave-assisted SPPS: Optimization of protocols and synthesis of difficult sequences

Precise microwave heating has emerged as a valuable addition to solid-phase peptide synthesis. New methods and protocols are required to utilize this potential and make it a reliable technique. Here, we describe a new semi-automated instrument for solid-phase peptide synthesis with microwave heating and report protocols for its reliable use. The instrument features a flow-through reaction vessel that is placed in the microwave instrument throughout the synthesis. Bubbling with N(2) gas during the microwave irradiation proved important for temperature control. Washing and Fmoc removal steps were automated, while activated Fmoc-amino acids were added manually. Several linkers and resins were evaluated and protocols for synthesis of difficult sequences were developed. These included on-resin reductive amination of BAL handles under microwave heating. We believe that the instrument, as well as the protocols for its use, will be useful tools for peptide chemists.