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

• DOI: 10.1016/j.microc.2022.107896
• Publication Date: 2022-08-28T15:18:27Z
• Authors: Anabel Torrente-L...; Jesús Hermosilla; Raquel Pérez-Robles; Antonio Salmerón-...; José Cabeza; Natalia Navas
• Keywords: Orbitrap

Nivolumab (Opdivo®) is a fully human immunoglobulin G4 isotype approved for the treatment of many cancers. It acts as an immune checkpoint inhibitor by blocking interaction between PD-1 (Programmed Cell Death Protein 1) – inhibitory receptor expressed on activated T cells- and its ligands, PD-L1 PD-L2. The quantification 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 UV MS data to full file requirement ICH-Q2(R1) develop stability (RP)UHPLC/UV-(HESI/Orbitrap™)MS method nivolumab medicinal A comparative study all figures merit or are shown discussed. results show linearity was similar two detectors established over range 4–45 μg/mL 1–45 (HESI/Orbitrap™)MS signals, respectively. sensitivity higher when signal (0.2 μg/mL) than with UV(2.0 μg/mL). However, provided better accuracy precision signal, which did not meet criteria robustness system suitability. spite this, plays crucial role methodology obtaining molecular weight profile 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 isoform 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.