biostudies-literatureLarson NRNIA NIH HHSNational Institutes of Health680-690https://www.ebi.ac.uk/biostudies/studies/S-EPMC9974558biostudies-literatureUnknown112(3)Liquid-liquid phase separation is a phenomenon within biology whereby proteins can separate into dense and more dilute phases with distinct properties. Three antibodies that undergo liquid-liquid phase separation were characterized in the protein-rich and protein-poor phases. In comparison to the protein-poor phase, the protein-rich phase demonstrates more blue-shift tryptophan emissions and red-shifted amide I absorbances. Large changes involving conformational isomerization around disulfide bonds were observed using Raman spectroscopy. Amide I and protein fluorescence differences between the phases persisted to temperatures above the critical temperature but ceased at the temperature at which aggregation occurred. In addition, large changes occurred in the structural organization of wateJournal of pharmaceutical sciencesConformational Changes and Drivers of Monoclonal Antibody Liquid-Liquid Phase Separation.PMC9974558R21 AG070466Forrest MLKalonia CKMiddaugh CREsfandiary RLarson NRCruz TAWei YfalseConformational Changes and Drivers of Monoclonal Antibody Liquid-Liquid Phase Separation.Liquid-liquid phase separation is a phenomenon within biology whereby proteins can separate into dense and more dilute phases with distinct properties. Three antibodies that undergo liquid-liquid phase separation were characterized in the protein-rich and protein-poor phases. In comparison to the protein-poor phase, the protein-rich phase demonstrates more blue-shift tryptophan emissions and red-shifted amide I absorbances. Large changes involving conformational isomerization around disulfide bonds were observed using Raman spectroscopy. Amide I and protein fluorescence differences between the phases persisted to temperatures above the critical temperature but ceased at the temperature at which aggregation occurred. In addition, large changes occurred in the structural organization of wate2023-01-01T00:00:00Z2023 Mar2025-04-03T23:50:04.179Z2025-04-03T23:50:04.179ZS-EPMC99745583630686210.1016/j.xphs.2022.10.017