RT Journal Article
T1 Combined use of UV and MS data for ICH Stability-Indication Method: Quantification and isoforms identification of intact nivolumab
A1 Torrente-Lopez, Anabel
A1 Hermosilla, Jesus
A1 Perez-Robles, Raquel
A1 Salmeron-Garcia, Antonio
A1 Cabeza, Jose
A1 Navas, Natalia
K1 UV
K1 MS data combined use
K1 Quantification
K1 Isoform profile identification
K1 Nivolumab
K1 LC-MS
K1 Size-exclusion chromatography
K1 Cell lung-cancer
K1 Monoclonal-antibody
K1 Mass-spectrometry
K1 Exchange chromatography
K1 Liquid-chromatography
K1 Validation
K1 Proteins
K1 Light
AB Nivolumab (Opdivo (R)) is a fully human immunoglobulin G4 isotype approved for the treatment of many cancers. It acts as an immune checkpoint inhibitor by blocking the interaction between PD-1 (Programmed Cell Death Protein 1) - an inhibitory receptor expressed on activated T cells- and its ligands, PD-L1 and PD-L2. The quantification of therapeutic proteins in their medicines and pharmaceutical preparations remains challenging because the protein content, a critical quality attribute, must be rigorously calculated using a validated stabilityindicating method, such as that indicated by the International Conference on Harmonization (ICH) quality guidelines, and this requires the analysis of the drug in the presence of its degraded products. In this work, we present an strategy based on the combined use of the UV and MS data to full file the requirement of the ICH-Q2 (R1) to develop and validated as stability indicated a (RP)UHPLC/UV-(HESI/OrbitrapTM)MS method for the quantification of nivolumab in medicinal products. A comparative study of all figures of merit of the method using UV or MS data are shown and discussed. The results show that linearity was similar for the two detectors and was established over a range of 4-45 mu g/mL and 1-45 mu g/mL for the UV and (HESI/OrbitrapTM)MS signals, respectively. The sensitivity of the method was higher when using the (HESI/OrbitrapTM)MS signal (0.2 mu g/mL) than with the UV(2.0 mu g/mL). However, the UV signal provided better accuracy and precision than the (HESI/ OrbitrapTM)MS signal, which did not meet the criteria for method robustness and system suitability. In spite of this, the MS signal plays a crucial role in this methodology by obtaining the molecular weight profile of the nivolumab isoforms, so enabling us to propose the glycans profile and detect structural modification due to degradation. The specificity of the method was evaluated by conducting forced degradation tests on samples of nivolumab in medicine form. The aim was to find out whether nivolumab suffers structural modifications when subject to stress. Structural modifications were detected by analysing the MS isoform profile, as changes of this kind promote new isoforms that are not chromatographically separated or detected by the UV signal. In this way, we demonstrated that the (RP)UHPLC/UV-(HESI/OrbitrapTM)MS method was capable of detecting nivolumab degradation, and was suitable for use in nivolumab stability studies. Thus, the protein content in the daily surplus of the Opdivo (R) medicine, stored either at room temperature (20 degrees C) or refrigerated at 4 degrees C, could be tracked for 15 days.
PB Elsevier
SN 0026-265X
YR 2022
FD 2022-08-17
LK http://hdl.handle.net/10668/22392
UL http://hdl.handle.net/10668/22392
LA en
NO Anabel Torrente-López, Jesús Hermosilla, Raquel Pérez-Robles, Antonio Salmerón-García, José Cabeza, Natalia Navas,Combined use of UV and MS data for ICH Stability-Indication Method: Quantification and isoforms identification of intact nivolumab,Microchemical Journal, Volume 182, 2022-08-17, 12
NO Anabel Torrente-Lopez ´ is currently receiving a FPU predoctoral grant (ref.: FPU18/03131) from the Ministry of Universities, Spain. JesúsHermosilla is currently benefiting from a research contract (P20_01029)from the Junta de Andalucía (Spain) and European Regional Development Funds. Raquel P´erez-Robles is currently granted a postdoctoralposition from the Junta de Andalucía, Spain.This study was funded by Project P20-01029 (I + D + i - Junta deAndalucía, Spain) and by Project B-FQM-308-UGR20 (Universidad deGranada, Proyectos I + D + i del Programa Operativo FEDER Andalucía2020) which means that it was also partially supported by EuropeanRegional Development Funds. Funding for open access charge: CBUA/Universidad de Granada.
DS RISalud
RD Apr 17, 2025