Development of attenuated total reflection infrared (ATR-IR) spectroscopic method is aimed for the high throughput rapid studies of large proteins solved in manufactured products. The fine-structural investigations – on atomic scale – will be extended by small angle X-ray scattering method providing the determination of protein-shapes. We intend to apply the combination of the two methods in the quality assurance processes. We set out the development of an innovative procedure for IR spectrum evaluation based on the variation of optical and sampling parameters. We have planed the establishment of the preliminary database of an IR spectrum library. We intend to study whether the change in the buffer system cause alterations in the IR spectra. We plan to follow the structural changes of proteins induced by different stress (treatments with ultrasound and gamma rays) presumably. We intend to reveal how the parameters of spectroscopic measurements affect the spectral signal to noise ratios, the output of the spectrum transformations and the results of structural estimations. By using the means of the two methods, we will study whether the spectroscopically revealed inner atomic structure, connected to the biological activity of the targeted protein, accompanies with changes in conformations related to the shape-alterations of whole proteins. In the first stage, model proteins such as lysozyme, trypsin, BSA will be studied, than the investigations will be extended towards pharmaceutical important products (e.g. monoclonal antibodies).

András Salgó, Attila Bóta, Szilveszter Gergely

Result_May 2020