RT Journal Article
T1 Comprehensive biophysical and functional study of ziv-aflibercept: characterization and forced degradation.
A1 Hermosilla, Jesus
A1 Perez-Robles, Raquel
A1 Salmeron-Garcia, Antonio
A1 Casares, Salvador
A1 Cabeza, Jose
A1 Bones, Jonathan
A1 Navas, Natalia
K1 Hypertonic Solutions
K1 Intravitreal Injections
K1 Light
K1 Receptors, Vascular Endothelial Growth Factor
K1 Recombinant Fusion Proteins
K1 Vascular Endothelial Growth Factor A
AB Aflibercept (AFL) is an Fc fusion protein used in the treatment of colorectal cancers and different ophthalmological diseases. There are two medicines in which AFL is the active substance: Zaltrap and Eylea, referred as ziv-AFL and AFL respectively. No proper accelerated degradation studies were published on either AFL or ziv-AFL. These studies are essential during research, development and manufacturing stages. Here, we characterized ziv-AFL and submitted it to different stress conditions: light, 60 °C, freeze-thaw cycles, changes in pH, high hypertonic solution and strong denaturing conditions. We used an array of techniques to detect aggregation (SE-HPLC/DAD and DLS), changes in secondary structure (Far-UV circular dichroism), changes in conformation or tertiary structure (Intrinsic tryptophan fluorescence) and alterations in functionality (ELISA). Results indicate that aggregation is common degradation pathway. Two different types of aggregates were detected: dimers and high molecular weight aggregates attributed to β-amyloid-like structures. Secondary structure was maintained in most of the stress tests, while conformation was altered by almost all the tests except for the freeze-thaw cycles. Functionality, evaluated by its immunochemical reaction with VEGF, was found to be stable but with decrease when exposed to light and with likely partial inactivation of the drug when pH was altered.
PB Nature Publishing Group
YR 2020
FD 2020-01-28
LK http://hdl.handle.net/10668/15109
UL http://hdl.handle.net/10668/15109
LA en
NO Hermosilla J, Pérez-Robles R, Salmerón-García A, Casares S, Cabeza J, Bones J, et al. Comprehensive biophysical and functional study of ziv-aflibercept: characterization and forced degradation. Sci Rep. 2020 Feb 14;10(1):2675.
NO This study was entirely funded by Project FIS: PI-17/00547 (Instituto Carlos III, Ministerio de Economía y Competitividad, Spain), which means that it was also partially supported by European Regional Development Funds (ERDF).
DS RISalud
RD Apr 7, 2025