Application of ATR-FTIR as a screening method for analysis of biopharmaceutical preparations containing trastuzumab

Abstract

In recent years, the number of therapeutic proteins introduced to the biopharmaceutical market has increased significantly, as medicine research and development have focused on protein-based therapies. Along with the introduction of biosimilar and biobetter medicines, the competitiveness of the biopharmaceutical market has grown considerably (Torres-Obreque et al., 2022). As a result, there is a global emergence of falsified biopharmaceuticals, especially monoclonal antibodies (mAbs), including trastuzumab, a humanized IgG1 mAb used as HER2-targeted therapy for the treatment of breast cancer (Maadi et al., 2021). In most of the seized falsified samples of medicines declared to contain mAbs (trastuzumab, rituximab, bevacizumab), no active pharmaceutical ingredient (API) was detected (Janvier et al, 2018). Therefore, suitable testing strategy and workflow should be defined in cases where falsification is suspected. Taking into account that mAbs are predominantly glycoproteins of high molecular weight (~ 150 kDa) with complex higher-order structure, their characterization can be rather challenging. Most of the analytical methods used for protein identification and characterization of mAbs (chromatography, electrophoresis, mass spectrometry and nuclear magnetic resonance) are time-consuming, expensive and require extensive sample preparation (Alhazmi et al., 2023). Therefore, implementation of fast, simple, and direct screening tools is of vast importance for the analytical quality control of these medicines. The use of vibrational spectroscopic techniques, particularly infrared (Hamla et al., 2022) for characterization of mAbs is drawing more attention since these are rapid and non-destructive methods. In this study, we demonstrate the potential use of Fourier transform infrared (FTIR) spectrophotometry, with attenuated total reflectance (ATR) for identification of trastuzumab in biopharmaceutical preparations.