Department of Pharmaceutical Sciences, School of Pharmacy, University of Maryland , Baltimore, Maryland 21201, United States.
Formulation Sciences, MedImmune , One MedImmune Way, Gaithersburg, Maryland 20878, United States.
Anal Chem. 2017 May 16;89(10):5494-5502. doi: 10.1021/acs.analchem.7b00464. Epub 2017 May 3.
Formulation stability is a critical attribute of any protein-based biopharmaceutical drug due to a protein's inherent tendency to aggregate. Advanced analytical techniques currently used for characterization of protein aggregates are prone to a number of limitations and usually require additional manipulations with the sample, such as dilution, separation, labeling, and use of special cuvettes. In the present work, we compared conventional techniques for the analysis of protein aggregates with a novel approach that employs the water proton transverse relaxation rate R(HO). We explored differences in the sensitivity of conventional techniques, size-exclusion chromatography (SEC), microflow imaging (MFI), and dynamic light scattering (DLS), and water NMR (wNMR) toward the presence of monoclonal antibody aggregates generated by different stresses. We demonstrate that wNMR outperformed SEC, DLS, and MFI in that it was most consistently sensitive to increases in both soluble and insoluble aggregates, including subvisible particles. The simplicity of wNMR, its sensitivity, and possibility of noninvasive measurements are unique advantages that would permit its application for more efficient and higher throughput optimization of protein formulations.
由于蛋白质固有的聚集倾向，制剂稳定性是任何基于蛋白质的生物制药的关键属性。目前用于蛋白质聚集物特性描述的先进分析技术容易受到许多限制，并且通常需要对样品进行额外的操作，例如稀释、分离、标记和使用特殊比色皿。在本工作中，我们将常规技术与一种新方法进行了比较，该方法采用水质子横向弛豫率 R(HO)。我们探讨了常规技术（SEC、MFI 和 DLS）与水 NMR（wNMR）之间的差异，以及它们对不同应激产生的单克隆抗体聚集物的敏感性。我们证明 wNMR 优于 SEC、DLS 和 MFI，因为它对可溶性和不溶性聚集物（包括亚可见颗粒）的增加最敏感。wNMR 的简单性、灵敏度和非侵入性测量的可能性是其独特的优势，这将使其能够更有效地应用于更高通量的蛋白质制剂优化。
