Direkte Affinitätsreinigung lang wirksamer PASylierter Proteine mit therapeutischem Anwendungspotential unter Nutzung von L‐Prolinamid zur milden Elution

Both insufficient plasma half-life (circulation for only few hours or less) and laborious downstream purification can be bottlenecks biological drug development. We report a novel strategy the efficient gentle affinity of pharmacologically relevant proteins modified by PASylation prolonged action in vivo . previously described antibodies specific Pro/Ala-rich sequences (PAS) covering range binding characteristics. Our approach relies on chromatography matrix functionalized with low-affinity PAS-specific antibody Fab fragment adsorption PASylated protein from macromolecular mixture. With complete absence hydrophobic/aromatic ionic groups PAS sequence epitope, is mediated Van der Waals contacts distinct hydrogen bonds only. Surprisingly, selective competitive elution achieved application highly soluble biologically inactive imino acid derivative L-prolinamide. Based but strongly dynamic biomolecular interaction, our procedure allows direct one-step cell extract culture supernatant while avoiding harsh conditions as they are often needed conventional chromatography.