20.500.12556/DKUM-98515 Application of NMR spectroscopy for studies of self-association and aggregation in protein formulations Protein self-association and aggregation are common phenomena that occur in various environments, including live cells, research samples, and during bioprocessing or storage of biopharmaceuticals. They may be a part of native biological function, or cause diseases, it can be an artefact in protein research, a concerning phenomenon in biopharmaceutical protein formulation, or a favourable opportunity to spontaneously concentrate proteins. The consequences of protein self-association and aggregation vary across different fields, and therefore may require somewhat different analytical approaches for their assessment and characterization. In this review we focus on types of aggregation and self-association which occur in protein formulations prepared for diverse purposes, where the aggregation itself is not the main functional feature of a protein. We aim to first highlight some major pathways of protein self-interaction and outline the terminology around the process of protein molecules clumping together, and the level of structural changes involved for each major pathway. We will briefly overview various analytical methods for characterising protein aggregation and self-association, and then consider the role of NMR, highlighting NMR parameters that are frequently used to gain insight into these processes. We focus on signal line broadening, chemical shift perturbation, diffusion coefficients, relaxation parameters and spatially selective NMR as the main approaches used to characterise various protein particles and the kinetics of their formation. Lastly, we discuss in more detail a few recent examples of NMR applications to study protein self-association and aggregation mainly in biopharmaceutical context, of how NMR measurables can assist in profiling of higher-order-structure, studies of protein-excipient interactions in formulation development, study of liquid-liquid phase separation and assessment of protein behaviour in complex mixtures. We also discuss low-field NMR methods that are being developed for in-line process monitoring as well as quality control. biopharmaceuticals protein aggregation prote ATPsdelf-association co-solvents excipients NMR biofarmacija proteini true false true Elsevier B.V. Angleški jezik Slovenski jezik https://www.sciencedirect.com/journal/progress-in-nuclear-magnetic-resonance-spectroscopy/vol/156/suppl/C © 2026 The Authors Članek v reviji 2026-06-17 16:01:30 2026-06-17 16:01:30 2026-06-18 03:25:30 0000-00-00 00:00:00 2026 0 0 28 str. article no.] 101611 Vol. 156/157 2026 0000-00-00 Zaloznikova Objavljeno NiDoloceno 2025-12-05 2026-05-22 2026-05-25 543.384 281382147 1873-3301 1-s2.0-S0079656526000178-main.pdf 1-s2.0-S0079656526000178-main.pdf 1 48BF26B86F07868C49C07C3B9A41FF9F 901bbbc215ad491a2beda7f8488f163efa4957310c96c49d6afd045305ab4814 57ae19d2-6a55-11f1-87c9-00155d580e34 https://dk.um.si/Dokument.php?lang=slv&id=207262 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0