2D NMR Analysis as a Sensitive Tool for Evaluating the Higher-Order Structural Integrity of Monoclonal Antibody against COVID-19
Francesca Cantini,
Emanuele Andreano,
Ida Paciello,
Veronica Ghini,
Francesco Berti,
Rino Rappuoli,
Lucia Banci
Affiliations
Francesca Cantini
Magnetic Resonance Center (CERM), Consorzio Interuniversitario Risonanze Magnetiche Metallo Proteine (CIRMMP), Department of Chemistry “Ugo Schiff”, University of Florence, 50019 Sesto Fiorentino, Italy
Magnetic Resonance Center (CERM), Consorzio Interuniversitario Risonanze Magnetiche Metallo Proteine (CIRMMP), Department of Chemistry “Ugo Schiff”, University of Florence, 50019 Sesto Fiorentino, Italy
Magnetic Resonance Center (CERM), Consorzio Interuniversitario Risonanze Magnetiche Metallo Proteine (CIRMMP), Department of Chemistry “Ugo Schiff”, University of Florence, 50019 Sesto Fiorentino, Italy
The higher-order structure (HOS) of protein therapeutics has been confirmed as a critical quality parameter. In this study, we compared 2D 1H-13C ALSOFAST-HMQC NMR spectra with immunochemical ELISA-based analysis to evaluate their sensitivity in assessing the HOS of a potent human monoclonal antibody (mAb) for the treatment of coronavirus disease 2019 (COVID-19). The study confirmed that the methyl region of the 2D 1H-13C NMR spectrum is sensitive to changes in the secondary and tertiary structure of the mAb, more than ELISA immunoassay. Because of its highly detailed level of characterization (i.e., many 1H-13C cross-peaks are used for statistical comparability), the NMR technique also provided a more informative outcome for the product characterization of biopharmaceuticals. This NMR approach represents a powerful tool in assessing the overall higher-order structural integrity of mAb as an alternative to conventional immunoassays.
