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

• Published in: Journal of magnetic resonance (1997) 2019-10, Vol.307, p.106581-106581, Article 106581
• Main Authors: Brinson, Robert G., Marino, John P.
• Format: Article
• Language: English
• Subjects: 2D NMR; Animals; Biologic; Biological Factors - therapeutic use; COIN-TACSY; COSY; Egg Proteins - chemistry; Fingerprinting; Higher order structure; Indicators and Reagents; Magnetic Resonance Spectroscopy; Muramidase - chemistry; Nuclear Magnetic Resonance, Biomolecular - methods; Peptide Mapping - methods; Protons; TOCSY
• ISSN: 1090-7807; 1096-0856
• DOI: 10.1016/j.jmr.2019.106581

[Display omitted] •2D NMR provides fingerprints of the higher-order-structure of protein therapeutics.•Fingerprint of 1Hn-1Hα correlations acquired with 2D J-correlated NMR experiments.•Selective TOCSY (TACSY) enables selective J-correlation and optimal sensitivity. The higher order structure (HOS) of protein therapeutics is essential for drug safety and efficacy and can be evaluated by two-dimensional (2D) nuclear magnetic resonance (NMR) spectroscopy at atomic resolution. 1Hn-15N amide correlated and 1H-13C methyl correlated NMR spectroscopies at natural isotopic abundance have been demonstrated as feasible on protein therapeutics as large as monoclonal antibodies and show great promise for use in establishing drug substance structural consistency across manufacturing changes and in comparing a biosimilar to an originator reference product. Spectral fingerprints from 1Hn-1Hα correlations acquired using 2D homonuclear proton-proton J-correlated NMR experiments provide a complementary approach for high-resolution assessment of the HOS of lower molecular weight (