Decoupling Protein Concentration and Aggregate Content Using Diffusion and Water NMR

IF 6.7 1区化学 Q1 CHEMISTRY, ANALYTICAL Analytical Chemistry Pub Date : 2024-06-29 DOI:10.1021/acs.analchem.3c05875

Mark I. Grimes, Matthew Cheeks, Jennifer Smith, Fabio Zurlo and Mick D. Mantle*,

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Abstract

Protein-based biopharmaceutical drugs, such as monoclonal antibodies, account for the majority of the best-selling drugs globally in recent years. For bioprocesses, key performance indicators are the concentration and aggregate level for the product being produced. In water NMR (wNMR), the use of the water transverse relaxation rate [R₂(¹H₂O)] has been previously used to determine protein concentration and aggregate level; however, it cannot be used to separate between them without using an additional technique. This work shows that it is possible to “decouple” these two key characteristics by recording the water diffusion coefficient [D(¹H₂O)] in conjunction with R₂(¹H₂O), even in the event of overlap in either D(¹H₂O) or R₂(¹H₂O). This method is demonstrated on three different systems, following appropriate D(¹H₂O) or R₂(¹H₂O) calibration data acquisition for a protein of interest. Our method highlights the potential use of benchtop NMR as an at-line process analytical technique.

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利用扩散和水核磁共振解耦蛋白质浓度和聚合体含量。

近年来，以蛋白质为基础的生物制药药物（如单克隆抗体）在全球最畅销的药物中占大多数。对于生物工艺而言，关键的性能指标是所生产产品的浓度和总量水平。在水核磁共振（wNMR）中，以前曾使用水的横向弛豫速率 [R2(1H2O)] 来确定蛋白质的浓度和聚集水平；但是，如果不使用额外的技术，就无法将它们区分开来。这项研究表明，通过记录水扩散系数[D(1H2O)]和 R2(1H2O)，即使 D(1H2O) 或 R2(1H2O)出现重叠，也能 "解耦 "这两个关键特征。我们在三个不同的系统上演示了这种方法，并在获取了相关蛋白质的适当 D(1H2O) 或 R2(1H2O) 校准数据后进行了验证。我们的方法强调了台式 NMR 作为在线过程分析技术的潜在用途。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Analytical Chemistry 化学-分析化学

CiteScore

12.10

自引率

12.20%

发文量

1949

审稿时长

1.4 months

期刊介绍： Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.