Physicochemical stability of reconstituted trastuzumab under freeze-thaw stress
Physicochemical stability of reconstituted trastuzumab under freeze-thaw stress
Afsaneh Farjami,1,*
1. Food and Drug Safety Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
Introduction: Trastuzumab is a recombinant humanized immunoglobulin G1 monoclonal antibody that is directed against human epidermal growthfactor receptor 2. Its action against CD20 leads to the death of CD20-positive cells, which is a desirable outcome. Because of the proteic nature of its structure, trastuzumab is vulnerable to a wide variety of chemical and physical degradation processes when subjected to a variety of various kinds of stress.
Methods: In this study, the stability of reconstitued trastuzumab was investigated when subjected to a number of different freeze-thaw cycles using a variety of orthogonal techniques, such as cation-exchange chromatography, size exclusion chromatography, attenuated total reflectance-Fourier infrared spectroscopy, and dynamic light scattering.
Results: According to the findings, reconstituted trastuzumab that had been kept in polyolefine bags remained stable after going through one cycle of freezing and thawing. There was no indication of either physical or chemical instability that was discovered. The hydrodynamic diameter of trastuzumab was found to be unaltered in the observations made. When compared to the profile of the control sample, the charge heterogeneity profile of the samples that were stressed did not change as a result of the freeze-thaw cycles. By using cation exchange chromatography, it was possible to determine that there was neither a new peak nor a reduction in the area under the curve. The protein's secondary structure did not undergo any changes as a result of the experiment.
Conclusion: In order to ensure the physicochemical stability of mAbs in addition to their biological stability, additional research is required to test the bioactivity of the stressed sample using an appropriate bioassay method. This is necessary in order to guarantee that mAbs will remain stable over time.