A deep dive into spin-labeled polysorbate’s interaction with therapeutic antibody using 2D NMR, EPR and MD simulations

Lebar, Blaž (avtor), Orehova, Marija Vjačeslavovna (avtor), Japelj, Boštjan (avtor), Šprager, Ernest (avtor), Podlipec, Rok (avtor), Knaflič, Tilen (avtor), Urbančič, Iztok (avtor), Knez, Benjamin (avtor), Zidar, Mitja (avtor), Cerar, Jure (avtor), Mravljak, Janez (avtor), Žula, Aleš (avtor), Arčon, Denis (avtor), Plavec, Janez (avtor), Pajk, Stane (avtor)

Polysorbates (PS) are widely used surfactants in biopharmaceutical formulations playing a crucial role in protecting proteins against mechanical stress and interface-induced damage. However, their susceptibility to degradation can compromise their function and lead to particle formation. Recent studies suggest that monoclonal antibodies (mAbs) may mitigate PS degradation catalyzed by histidine chloride buffer, indicating the presence of protein-PS interactions. In this study, we investigated these interactions using NMR, starting with $^1$H T$_2$ CPMG filter experiments and methyl fingerprinting, which failed to detect interactions. To enhance sensitivity, we synthesized spin-labeled PS (SLPS), enabling paramagnetic relaxation enhancement (PRE) NMR experiments, specifically amide fingerprinting, which successfully revealed interactions. Complementary electron paramagnetic resonance (EPR) measurements of SLPS and mAb also detected interactions, but only in the presence of sucrose, underscoring their weak and transient nature. Additionally, molecular dynamics simulations identified potential interaction hotspots on the antibody structure, providing mechanistic insights into these interactions.

• Jezik: Angleški jezik
• Ključne besede: monoclonal antibody, interaction, polysorbate, nuclear magnetic resonance, electron paramagnetic resonance, molecular dynamics
• Vrsta gradiva: Članek v reviji
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: FFA - Fakulteta za farmacijo
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2025
• Št. strani: 9 str.
• Številčenje: Vol. 217, art. 114892
• PID: 20.500.12556/RUL-182732
• UDK: 577:615.32
• ISSN pri članku: 1873-3441
• DOI: 10.1016/j.ejpb.2025.114892
• COBISS.SI-ID: 255171075
• Datum objave v RUL: 21.05.2026

Gradivo je del revije

• Naslov: European journal of pharmaceutics and biopharmaceutics
• Založnik: Elsevier
• ISSN: 1873-3441
• COBISS.SI-ID: 23171845

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: monoklonsko protitelo, interakcija, polisorbat, jedrska magnetna resonanca, elektronska paramagnetna resonanca, molekularna dinamika, farmacevtska kemija, biokemija, zdravila

Projekti

• Financer: Novartis Pharmaceutical Manufacturing LLC

• Financer: ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
• Številka projekta: P1-0208
• Naslov: Farmacevtska kemija: načrtovanje, sinteza in vrednotenje učinkovin