Department of Pharmaceutical Sciences, School of Pharmacy , University of Maryland , Baltimore , Maryland 21201 , United States.
Anal Chem. 2019 Nov 5;91(21):13538-13546. doi: 10.1021/acs.analchem.9b02622. Epub 2019 Oct 8.
Continuous manufacturing of biologics is one of the priorities of the biopharmaceutical industry. However, its widespread implementation is hampered by a lack of noninvasive/nondestructive process analytical technology (PAT) systems capable of real-time in-line monitoring of product flow parameters, such as concentration and/or aggregate content. We have previously demonstrated that, under nonflow conditions, the water proton transverse relaxation rate, (HO), is sensitive to protein concentration and aggregate content in biopharmaceutical formulations. In the present work, we explored the potential of water proton NMR under flow conditions (flow-NMR) to use (HO) as a quantitative indicator of protein concentration variations and aggregate levels in the process flow. We show that, under flow conditions, (HO) is sensitive to rather small changes in protein concentration (<1 mg/mL) and is capable to detect variations in the aggregate content of <1%. Our findings suggest that flow-NMR could be advantageously used as a real-time in-line noninvasive PAT for continuous biomanufacturing.
生物制药行业的重点之一是连续制造生物制剂。然而，由于缺乏能够实时在线监测产品流动参数（如浓度和/或聚集物含量）的非侵入式/非破坏性过程分析技术（PAT）系统，其广泛应用受到阻碍。我们之前已经证明，在非流动条件下，水质子横向弛豫率（HO）对生物制药制剂中的蛋白质浓度和聚集物含量敏感。在本工作中，我们探索了流动条件下水质子 NMR（flow-NMR）的潜力，以将（HO）用作蛋白质浓度变化和过程流中聚集物水平的定量指标。我们表明，在流动条件下，（HO）对蛋白质浓度的微小变化（<1mg/mL）敏感，并且能够检测到<1%的聚集物含量变化。我们的研究结果表明，flow-NMR 可以作为连续生物制造的实时在线非侵入式 PAT 优势使用。
