Benefits of a Mixed-Mode Stationary Phase to Address the Challenging Purification of an Industrially Relevant Peptide: A Proof-of-Concept Study

Peptides are a class of biomolecules with great potential from the therapeutic point view, because their unique biological properties. Industrially, production stategies adopted produce both target peptide and series impurities that must be removed. Preparative chromatography is technique choice for large-scale purification biomolecules, generally performed in reversed-phase mode, using hydrophobic adsorbents (e.g., C8 stationary phases). A promising innovative alternative represented by mixed-mode columns, which bear two different ligands on particle surface, exploiting retention mechanisms to improve separation. This work represents proof-of-concept study focused comparison adsorbent one (bearing groups charged ones) crude mixture. Thanks more-favourable thermodynamics, it was found that, when collecting whole peak excluding fractions tail, column led an increase recovery roughly +15%, together slight improvement purity at same time, respect traditional column. In addition, if peak, including collected, performance columns similar terms recovery, but pepetide elutes as narrower mixed mode. leads collection pool showing much-higher concentration lower solvent volumes needed, beneficial achievement targeting more sustainable processes. These results very advantageous industrial viewpoint, they also involve decrease amount contained reprocessed again satisfy requirements.