Comprehensive Assessment of Protein and Excipient Stability in Biopharmaceutical Formulations Using (1)H NMR Spectroscopy.
Comprehensive Assessment of Protein and Excipient Stability in Biopharmaceutical Formulations Using (1)H NMR Spectroscopy.
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DOI：
10.1021/acsptsci.0c00188
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发表时间：
2021-02-12
影响因子：
--
通讯作者：
Golovanov AP
中科院分区：
文献类型：
--
作者：
Bramham JE;Podmore A;Davies SA;Golovanov AP
生物药物蛋白质是治疗一系列疾病的重要药物疗法。蛋白质，如抗体（Ab）和肽，易于化学和物理降解，特别是在目前寻求皮下注射的高浓度下，因此必须优化制剂条件，包括缓冲液和赋形剂，以最大限度地减少此类不稳定性。因此，生物制药制剂的蛋白质和小分子含量及其稳定性对于治疗的成功至关重要。然而，评估蛋白质和小分子稳定性的所有方面目前需要大量的分析技术，其中大多数涉及样品稀释或其他操作，这些操作本身可能会扭曲样品行为。在这里，我们展示了应用1H核磁共振（NMR）光谱研究蛋白质和小分子的含量和稳定性原位高浓度（100 mg/mL）抗体制剂。我们发现，蛋白质降解（聚集或碎片）可以检测到的变化，在1D 1H NMR信号强度，而表观弛豫速率是特别敏感的抗体片段。同时，弛豫过滤光谱揭示了小分子组分（如赋形剂）的存在和降解，以及一般溶液性质（如pH值）的变化。因此，1H NMR光谱可以提供生物制药制剂含量和稳定性的整体概述，提供降解的初步表征，并作为分类步骤，以指导进一步的分析技术。
Biopharmaceutical proteins are important drug therapies in the treatment of a range of diseases. Proteins, such as antibodies (Abs) and peptides, are prone to chemical and physical degradation, particularly at the high concentrations currently sought for subcutaneous injections, and so formulation conditions, including buffers and excipients, must be optimized to minimize such instabilities. Therefore, both the protein and small molecule content of biopharmaceutical formulations and their stability are critical to a treatment’s success. However, assessing all aspects of protein and small molecule stability currently requires a large number of analytical techniques, most of which involve sample dilution or other manipulations which may themselves distort sample behavior. Here, we demonstrate the application of 1H nuclear magnetic resonance (NMR) spectroscopy to study both protein and small molecule content and stability in situ in high-concentration (100 mg/mL) Ab formulations. We show that protein degradation (aggregation or fragmentation) can be detected as changes in 1D 1H NMR signal intensity, while apparent relaxation rates are specifically sensitive to Ab fragmentation. Simultaneously, relaxation-filtered spectra reveal the presence and degradation of small molecule components such as excipients, as well as changes in general solution properties, such as pH. 1H NMR spectroscopy can thus provide a holistic overview of biopharmaceutical formulation content and stability, providing a preliminary characterization of degradation and acting as a triaging step to guide further analytical techniques.
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影响因子：
3.7
作者：
Dion, Michelle Z.;Leiske, Danielle;Salisbury, Cleo M.
通讯作者：
Salisbury, Cleo M.
DOI：
10.1007/978-3-030-11751-1_6
发表时间：
2019-01-01
期刊：
SCIENCE AND REGULATIONS OF NATURALLY DERIVED COMPLEX DRUGS
影响因子：
--
作者：
Keire, David A.
通讯作者：
Keire, David A.
影响因子：
4.8
作者：
Joubert, Marisa K.;Luo, Quanzhou;Narhi, Linda O.
通讯作者：
Narhi, Linda O.
影响因子：
5.3
作者：
Kheddo P;Cliff MJ;Uddin S;van der Walle CF;Golovanov AP
通讯作者：
Golovanov AP
影响因子：
3.7
作者：
Nishi, Hirotaka;Miyajima, Makoto;Fukui, Kiichi
通讯作者：
Fukui, Kiichi