A multifaceted approach to understanding protein-buffer interactions in biopharmaceuticals

Lebar, Blaž (avtor), Orehova, Maria (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)

The excipient selection process plays a crucial role in biopharmaceutical formulation development to ensure the long-term stability of the drug product. Though there are numerous options approved by regulatory authorities, only a subset is commonly utilized. Previous research has proposed various stabilization mechanisms, including protein-excipient interactions. However, identifying these interactions remains challenging due to their weak and transient nature. In this study, we present a comprehensive approach to identify such interactions. Using the $^1$H T$_2$ CPMG (Carr-Purcel-Meiboom-Gill) filter experiment we identified interactions of rituximab with certain buffers and amino acids, shedding light on its Fc fragment instability that manifested during the enzymatic cleavage of the antibody. Moreover, chemometric analyses of 2D NMR fingerprints revealed interactions of selected excipients with antibody fragments. Furthermore, molecular dynamics simulations revealed potential interacting hotspots without NMR spectra assignment. Our results highlight the importance of an orthogonal method approach to uncovering these critical interactions, advancing our understanding of excipient stabilization mechanisms and rational formulation design in biopharmaceutics.

• Jezik: Angleški jezik
• Ključne besede: buffer, interaction, methyl fingerprinting, molecular dynamics, monoclonal antibody, nuclear magnetic resonance
• 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: 12 str.
• Številčenje: Vol. 206, art. 114582
• PID: 20.500.12556/RUL-165181
• UDK: 54:615
• ISSN pri članku: 0939-6411
• DOI: 10.1016/j.ejpb.2024.114582
• COBISS.SI-ID: 216273923
• Datum objave v RUL: 26.11.2024

Gradivo je del revije

• Naslov: European journal of pharmaceutics and biopharmaceutics
• Skrajšan naslov: Eur. j. pharm. biopharm.
• Založnik: Elsevier
• ISSN: 0939-6411
• COBISS.SI-ID: 13176839

Licence

• Licenca: CC BY-NC 4.0, Creative Commons Priznanje avtorstva-Nekomercialno 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by-nc/4.0/deed.sl
• Opis: Licenca Creative Commons, ki prepoveduje komercialno uporabo, vendar uporabniki ne rabijo upravljati materialnih avtorskih pravic na izpeljanih delih z enako licenco.

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: medpomnilnik, interakcija, metilni prstni odtisi, molekularna dinamika, monoklonsko protitelo, jedrska magnetna resonanca, pomožne snovi, zdravila

Projekti

• Financer: Drugi - Drug financer ali več financerjev
• Program financ.: 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