Department of Pharmaceutical Sciences , School of Pharmacy, University of Maryland , Baltimore , Maryland 21201 , United States.
Dosage Form Design and Development , MedImmune , One MedImmune Way , Gaithersburg , Maryland 20878 , United States.
Anal Chem. 2019 Mar 19;91(6):4107-4115. doi: 10.1021/acs.analchem.8b05733. Epub 2019 Feb 25.
Water proton transverse relaxation rate R(HO) measurements by NMR stand out as a powerful noninvasive tool to detect protein aggregates, including subvisible particles in biopharmaceutical formulations. To understand the applicability of water proton NMR ( wNMR), we studied the response and sensitivity of wNMR to the aggregates of a monoclonal antibody (mAb) within a wide size range at different aggregate levels, for three different physical stresses: freeze-thaw cycling, heating, and agitation. We compared the sensitivity and response of wNMR with those observed by conventional techniques of size exclusion chromatography (SEC), microflow imaging (MFI), and dynamic light scattering (DLS). Our findings showed that wNMR detects mAb aggregates within wide aggregate levels and in a wide range of aggregate sizes. wNMR was sensitive to an increase in soluble protein aggregates in the range of <1.0%. In most cases, wNMR demonstrated linear response toward the aggregate fraction. Nonlinearity of such response potentially points to the presence of larger size aggregates that possibly rearrange and/or dissociate upon dilution. The results demonstrate the potential of wNMR as a quantitative and noninvasive analytical tool for characterizing protein aggregates in biopharmaceutical formulations.
水质子横向弛豫率 R(HO) 的测量通过 NMR 成为一种强大的非侵入性工具，可用于检测蛋白质聚集体，包括生物制药制剂中的亚可见颗粒。为了了解水质子 NMR（wNMR）的适用性，我们研究了 wNMR 对单克隆抗体（mAb）聚集体的响应和敏感性，该聚集体在不同的聚集体水平和广泛的大小范围内，经历了三种不同的物理应力：冻融循环、加热和搅拌。我们将 wNMR 的灵敏度和响应与尺寸排阻色谱（SEC）、微流成像（MFI）和动态光散射（DLS）等常规技术观察到的结果进行了比较。我们的研究结果表明，wNMR 可以在广泛的聚集体水平和广泛的聚集体大小范围内检测到 mAb 聚集体。wNMR 对可溶性蛋白质聚集体增加<1.0%的范围敏感。在大多数情况下，wNMR 对聚集体分数表现出线性响应。这种响应的非线性可能表明存在更大尺寸的聚集体，这些聚集体可能在稀释时重新排列和/或解离。结果表明，wNMR 有潜力成为生物制药制剂中蛋白质聚集体的定量和非侵入性分析工具。
