Author
Taiji Oyama, Sales Division, JASCO Corporation Ken-ichi Akao and Satoko Suzuki, Applicative solution lab, JASCO Corporation
Keywords
Protein, Monoclonal antibody, Secondary structure estimation, BeStSel, β-sheet
Abstract
Circular dichroism spectroscopy is widely used to estimate the conformation of polymers, and higher-order structures of proteins and nucleic acids. In particular, CD spectra in the far-ultraviolet region contain an abundant information on the secondary structure of proteins. Therefore, far-UV CD spectroscopy has long been used for structure and dynamics research of biomolecules in academia and is now widely used for research and development and quality control of pharmaceuticals as the market for protein and antibody drugs expands. In 2015, Beta Structure Selection (BeStSel) was developed by Kardos and Micsonai et al. BeStSel can accurately estimate the secondary structure of proteins, including β-sheet-rich proteins, by taking into account the twisting between β-strands. The BeStSel algorithm is integrated into Spectrum Manager Ver. 2.5 through a corroboration between Eötvös Loránd University and JASCO Corporation and developed as JWBeStSel-532. JWBeStSel-532 can provide not only an offline analysis environment but also seamless analysis platform from spectrum measurement to secondary structure estimation using BeStSel. The combination of J-1500, the ultra-short path-length cell, and BeStSel enables the estimation of the secondary structure of antibody drugs in the formulation state. In addition, the combination of BeStSel and HTCD Plus, which allows fully automated acquisition of CD, fluorescence, and absorption spectra, provides a comprehensive evaluation of the stability of the higher-order structure of antibody drugs.