Institute for Bioscience and Biotechnology Research, National Institute of Standards and Technology and the University of Maryland, 9600 Gudelsky Drive, Rockville, MD 20850, United States.
Institute for Bioscience and Biotechnology Research, National Institute of Standards and Technology and the University of Maryland, 9600 Gudelsky Drive, Rockville, MD 20850, United States.
J Magn Reson. 2019 Oct;307:106581. doi: 10.1016/j.jmr.2019.106581. Epub 2019 Aug 20.
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. Hn-N amide correlated and H-C 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 Hn-Hα 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 (<25 kDa) protein therapeutics. Here, we evaluate different pulse sequences (COSY, TOCSY and TACSY) used to generate proton-proton J-correlated NMR spectral fingerprints and appraise the performance of each method for application to protein therapeutic HOS assessment and comparability.
蛋白质治疗药物的高级结构（HOS）对于药物安全性和疗效至关重要，可以通过二维（2D）核磁共振（NMR）光谱技术在原子分辨率水平进行评估。已经证明，在天然同位素丰度下，Hn-N 酰胺相关和 H-C 甲基相关 NMR 光谱技术在单克隆抗体等大型蛋白质治疗药物上是可行的，并且在建立药物物质结构一致性方面具有很大的应用潜力，可用于比较生物类似药与原创参考产品。使用二维同核质子质子 J 相关 NMR 实验获得的 Hn-Hα 相关谱指纹提供了一种用于低分子量（<25 kDa）蛋白质治疗药物高级结构高分辨率评估的互补方法。在这里，我们评估了用于生成质子质子 J 相关 NMR 谱指纹的不同脉冲序列（COSY、TOCSY 和 TACSY），并评估了每种方法在蛋白质治疗药物高级结构评估和可比性方面的应用性能。
