In recent years, biological drugs have become the primary focus of research and development in biopharmaceuticals. Modern biological drugs are produced using recombinant technology in biological systems such as bacteria, yeast, or mammalian cells. These drugs are predominantly proteins, and due to their sensitivity to external conditions, they are often dispensed in liquid form. The stability of the active pharmaceutical ingredient in liquid form is affected by various excipients, including sugars and buffers. Sugars ensure long-term stability and protect proteins during cryogenic freezing and lyophilization, while buffers maintain an appropriate pH value and protect protein particles from degradation.
The standard practice for analyzing excipients in biopharmaceutical production involves the use of high-performance liquid chromatography (HPLC). This thesis evaluates the potential of Fourier-transform infrared spectroscopy (FT-IR) to provide comparable results in the analysis of various excipients. The basic concepts of HPLC and FT-IR techniques are presented, particularly with a modern MIRA analyzer, highlighting their main advantages, disadvantages, areas of application, and the materials and methods employed.
Two experiments were conducted to evaluate the impact of different analytical approaches on the development process of biological drugs. The first experiment involved the analysis of sugars (sucrose, trehalose, sorbitol, mannitol) at various concentrations using both methods. The second experiment focused on comparing the accuracy and reliability of the FT-IR and HPLC methods in the analysis of buffers at different concentration ranges (citrate, histidine, acetate). The results indicated that both methods offer comparable accuracy and reliability; however, sample preparation for HPLC is more time-consuming due to additional dilutions, which increases the potential for pipetting errors. Based on the obtained results, it was concluded that FT-IR enables faster quantitative and qualitative analysis compared to the HPLC method.
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