Combined use of UV and MS data for ICH Stability-Indication Method: Quantification and isoforms identification of intact nivolumab

• Combined used of UV and MS data allow for method validation as ICH stability indicating. signal showed clearly better parameters validation. HRMS was crucial to assure specificity. (RP)UHPLC/UV-(HESI/Orbitrap™)MS nivolumab quantification is shown the first time. Nivolumab isoform profile identified by mass spectrometry. (Opdivo®) a fully human immunoglobulin G4 isotype approved treatment many cancers. It acts an immune checkpoint inhibitor blocking interaction between PD-1 (Programmed Cell Death Protein 1) – inhibitory receptor expressed on activated T cells- its ligands, PD-L1 PD-L2. The therapeutic proteins in their medicines pharmaceutical preparations remains challenging because protein content, critical quality attribute, must be rigorously calculated using validated stability-indicating method, such that indicated International Conference Harmonization (ICH) guidelines, this requires analysis drug presence degraded products. In work, we present strategy based combined use full file requirement ICH-Q2(R1) develop medicinal A comparative study all figures merit or are discussed. results show linearity similar two detectors established over range 4–45 μg/mL 1–45 (HESI/Orbitrap™)MS signals, respectively. sensitivity higher when (0.2 μg/mL) than with UV(2.0 μg/mL). However, provided accuracy precision signal, which did not meet criteria robustness system suitability. spite this, plays role methodology obtaining molecular weight isoforms, so enabling us propose glycans detect structural modification due degradation. specificity evaluated conducting forced degradation tests samples medicine form. aim find out whether suffers modifications subject stress. Structural were detected analysing profile, changes kind promote new isoforms chromatographically separated signal. way, demonstrated capable detecting degradation, suitable studies. Thus, content daily surplus Opdivo® medicine, stored either at room temperature (20 °C) refrigerated 4 °C, could tracked 15 days.