3PC-065 Impact of light stress on the isoform profile of nivolumab (Opdivo) in opened vials estimated by (RP)UHPLC-UV-(HESI/Orbitrap)-MS

Background and importance Nivolumab (Opdivo) is a human IgG4 monoclonal antibody from the group of immunomodulators which bind to the programmed death receptor 1 (PD-1). As a complex protein, physical aggregation and chemical degradation can occur throughout its life, and even modest environmental stresses can cause extensive damage.1 As indicated in its technical report,2 the unopened vials can be stored at a controlled room temperature up to 25°C with room light for up to 48 hours. Aim and objectives To assess the impact on the isoform profile of nivolumab 10 mg/mL (Opdivo) promoted by exposure to light in its own opened vial at a controlled temperature of 25°C to evaluate likely risks from unintentional mishandling in real hospital conditions. Material and methods Nivolumab (Opdivo, 10 mg/mL) was placed in an accelerated stress test chamber to simulate sunlight (Solarbox 3000e RH, Cofomegra, Milan, Italy) for 24 hours at 25°C. Irradiation was set at 250 W/m2, between 320 and 800 nm.3 A validated reverse phase ultra high resolution liquid chromatography coupled to high resolution mass spectrometry and exact mass ((RP)UHPLC-UV-(HESI/Orbitrap)-MS) method was used to analyse intact nivolumab. UV-chromatograms and total ion chromatograms (TICs) were recorded and the deconvoluted mass spectra gave the nivolumab mass isoform profile. Results UV chromatograms and TICs suggested no degradation products after light exposure. However, isoform profiles clearly showed changes in the light submitted nivolumab samples. An important increase in the number of isoforms even with changes in their masses, including the main isoform, was detected. Conclusion and relevance Exposure to light may cause modifications in the nivolumab isoform profile which suggests protein degradation. This work shows the importance of protecting opened vials of the medicine Opdivo from light (and by extension, bags for infusion) when they are at room temperature (up to 25°C). References and/or acknowledgements Nejadnik MR, et al. J Pharm Sci 2018;107:2013–19. Nivolumab Technical Report. https://www.ema.europa.eu/en/documents/product-information/opdivo-epar-product-information_en.pdf Scientific discussion ICH Q1B photostability testing of new active substances and medicinal products. European Medicines Agency (EMEA) 1998 https://www.ema.europa.eu/en/documents/scientific-guideline/ich-q-1-b-photostability-testing-new-active-substances-medicinal-products-step-5_en.pdf Funded by project FIS: PI-17/00547 (Instituto Carlos III, Spain), which means that it was also partially supported by European Regional Development Funds. AT-L is currently receiving an FPU predoctoral grant (reference FPU18/03131) from the Ministry of Universities, Spain. Conflict of interest No conflict of interest