Torrente-Lopez, AnabelHermosilla, JesusSalmeron-Garcia, AntonioCabeza, JoseNavas, Natalia2023-05-032023-05-032022-03-17Torrente-López A, Hermosilla J, Salmerón-García A, Cabeza J, Navas N. Comprehensive Analysis of Nivolumab, A Therapeutic Anti-Pd-1 Monoclonal Antibody: Impact of Handling and Stress. Pharmaceutics. 2022 Mar 23;14(4):6921999-4923http://hdl.handle.net/10668/21547Nivolumab, formulated in the medicine Opdivo® (10 mg/mL), is a therapeutic monoclonal antibody (mAb) used in the treatment of different types of cancer. Currently, there is insufficient knowledge about the behaviour of this protein with regards to the risk associated with its routine handling or unintentional mishandling, or when subjected to stress conditions in hospitals. These conditions can be simulated in forced degradation studies, which provide an in-depth understanding of the biophysical and biochemical properties of mAbs. In this study, we carried out a physicochemical and functional characterisation of nivolumab, which was subjected to various stress conditions: heat, freeze/thaw cycles, agitation, light exposure and high hypertonic solution. We used a wide range of analytical techniques: Far-UV CD, IT-FS, DLS, SE/UHPLC(UV)-[Native]MS, and ELISA. The results show that exposure to light was the stress test with the greatest impact on the samples, revelling the formation of non-natural dimers and a different isoform profile. In addition, nivolumab (Opdivo®) demonstrated stability up to 60 °C (1 h). As regards functionality all the nivolumab (Opdivo®) stressed samples were found to be stable except for those subjected to light and agitation, and to a lesser extent, those subjected to FTC 5 and NaCl stresses.enAttribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/UHPLC-MScomprehensive analytical characterisationforced degradationisoform profilenivolumabNivolumabAntibodies, MonoclonalSodium ChlorideChromatography, High Pressure LiquidExercise TestNeoplasmsProtein IsoformsEnzyme-Linked Immunosorbent AssayHospitalsresearch article35456527open accessAnticuerpos monoclonalesCloruro de sodioCromatografía líquida de alta presiónEnsayo de inmunoadsorción enzimáticaHospitalesIsoformas de proteínasNeoplasiasNivolumabPrueba de esfuerzo10.3390/pharmaceutics14040692PMC9025134https://www.mdpi.com/1999-4923/14/4/692/pdf?version=1648033999https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9025134/pdf