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2D J-correlated proton NMR experiments for structural fingerprinting of biotherapeutics



Robert G. Brinson, John P. Marino


The higher order structure (HOS) of biotherapeutics is essential for drug safety and efficacy and can be evaluated by nuclear magnetic resonance (NMR) spectroscopy at atomic resolution. NMR spectral fingerprinting techniques based on 2D heteronuclear correlations of the amide or methyl regions of the protein spectrum have been demonstrated for protein biologics, including monoclonal antibodies (mAbs), and have applications in establishing drug substance structural consistency across manufacturing changes and in comparing a biosimilar to an originator reference product. Here, we evaluate an alternative approach to generating spectral maps for comparability which is based on 1HN-H correlations that are acquired using 2D homonuclear proton-proton J-correlated NMR experiments. We compare the performance of four different experimental approaches to generate these maps and appraise the utility of each for use as tools for HOS assessment and comparability.
Journal of Magnetic Resonance


proton NMR, fingerprinting, protein, peptide, COSY, TOCSY, COIN-TACSY, higher order structure


Brinson, R. and Marino, J. (2019), 2D J-correlated proton NMR experiments for structural fingerprinting of biotherapeutics, Journal of Magnetic Resonance, [online], (Accessed July 20, 2024)


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Created October 1, 2019, Updated February 13, 2020