Flow-based Peptide and Protein Synthesis

The field of biopharmaceuticals is rapidly expanding, requiring new methods for the on-demand production of chemically modified peptides and proteins. Chemical synthesis depends on efficient amide bond formation, with solid-phase peptide synthesis (SPPS) remaining the standard method for producing medium- to long-length peptides. However, yields often depend on the specific sequence, as peptide aggregation on the solid support can cause incomplete couplings in “difficult sequences.” Limited analytical tools have long hindered systematic study of this phenomenon. Flow-SPPS enables rapid synthesis and in-line analytics, offering insight into sequence-dependent aggregation. Aggregation phenomena are analyzed in detail, and novel computational, technological, and synthetic approaches are presented to minimize sequence dependence in SPPS.