Biosimilar medicines represent a more affordable alternative to original biological drugs, which have gained increasing importance in recent years and play a significant role in the treatment of complex diseases such as cancer, rheumatoid arthritis, Crohn's disease, psoriasis, and Alzheimer's disease. One of the important therapeutic targets of monoclonal antibodies is the TNF-α factor. By inhibiting it, we contribute to controlling inflammatory processes in numerous autoimmune diseases.
Protein structures are sensitive even to small changes in temperature, pH, osmolality, mechanical stress, the presence of oxygen, light, and other factors. The presence of moisture affects the physical, chemical, and microbiological stability of proteins, therefore, with lyophilization, which reduces moisture content, we improve the stability of biological medicines and extend their shelf life.
In this master’s thesis, we investigated the stability of a selected protein-based active pharmaceutical ingredient, which was exposed to various temperatures and relative humidity levels. We measured the reconstitution time, pH, osmolality, water content using Karl Fischer coulometric titration with an oven, and analysed protein purity using gel permeation chromatography. We found that the selected protein is stable under elevated temperature and humidity conditions. No significant differences in protein purity were observed between the different conditions. The aim of the thesis was also to compare two methods for measuring moisture - the gold standard Karl Fischer titration and a method for measuring headspace moisture using tunable diode laser absorption spectroscopy. We demonstrated that the two moisture measurement methods are comparable; however, due to the lack of sorption isotherm data for moisture for the selected pharmaceutical product, we cannot yet confirm the applicability or interchangeability of the method. We showed that, by using a non-destructive method for measuring moisture in the headspace, it is possible to obtain a rapid estimate of moisture content, which could potentially have a wider range of applications in the future and perhaps even replace Karl Fischer titration.
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