FT-IR versus EC-QCL spectroscopy for biopharmaceutical quality assessment with focus on insulin—total protein assay and secondary structure analysis using attenuated total reflection
Autor: | Sven Delbeck, H. Michael Heise |
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Jazyk: | angličtina |
Rok vydání: | 2020 |
Předmět: |
Infrared spectroscopy
030209 endocrinology & metabolism 01 natural sciences Biochemistry Protein Structure Secondary Analytical Chemistry Biopharmaceuticals 010309 optics 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Secondary structure analysis 0103 physical sciences Spectroscopy Fourier Transform Infrared Animals Humans Hypoglycemic Agents Insulin Fourier transform infrared spectroscopy Protein secondary structure Active ingredient Biological Products Chromatography Silver halide Insulin stability Quality control Proteins FT-IR-ATR spectroscopy Biopharmaceutical chemistry Attenuated total reflection Chemical stability Insulin fibrils Research Paper |
Zdroj: | Analytical and Bioanalytical Chemistry |
ISSN: | 1618-2650 1618-2642 |
Popis: | For the quality control of biopharmaceutical products, which contain proteins as the most important active ingredients, shelf life may be limited due to inappropriate storage conditions or mechanical stress. For insulins as representatives of life-saving pharmaceuticals, analytical methods are needed, which are providing additional information than obtained by assays for total protein quantification. Despite sophisticated formulations, the chemical stability may be challenged by temperatures deviating from recommended conditions or shear rate exposure under storage, leading to misfolding, nucleation, and subsequent fibril formation, accompanied by a decrease in bioactivity. A reliable method for insulin quantification and determination of secondary structure changes has been developed by attenuated total reflection (ATR) Fourier-transform infrared spectroscopy of insulin formulations by a silver halide fiber-coupled diamond probe with subsequent dry-film preparation. A special emphasis has been placed on the protein amide I band evaluation, for which spectral band analysis provides unique information on secondary structure fractions for intact and misfolded insulins. Quantitative measurements are possible down to concentrations of less than 0.5 mg/ml, whereas the dry-film preparation delivers high signal-to-noise ratios due to the prior water evaporation, thus allowing a reliable determination of secondary structure information. |
Databáze: | OpenAIRE |
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