Raman spectroscopy for in situ, real time monitoring of protein aggregation in lyophilized biotherapeutic products
Raman spectroscopy for in situ, real time monitoring of protein aggregation in lyophilized biotherapeutic products
复制标题
DOI：
10.1016/j.ijbiomac.2021.02.214
复制
发表时间：
2021-03-09
影响因子：
8.2
通讯作者：
Rathore, Anurag S.
中科院分区：
文献类型：
--
作者：
Nitika, Nitika;Chhabra, Hemlata;Rathore, Anurag S.
生物治疗产品的质量对于确保患者安全至关重要。人们非常需要能够在护理时快速评估治疗产品质量的分析工具。在本文中，我们应用基于化学计量学的拉曼光谱分析来定量预测冻干生物治疗产品中的蛋白质聚集。两种市售治疗蛋白促红细胞生成素（EPO）和人生长激素（HGH）已被用来证明所提出方法的适用性。通过尺寸排阻色谱法以及使用 785 nm 波长激光的拉曼光谱来监测热诱导的蛋白质聚集。使用偏最小二乘 (PLS) 回归分析拉曼光谱并创建用于定量测定聚集体的模型。 EPO 和 HGH 的性能均令人满意，R-2 分别为 0.91 和 0.94，交叉验证相关系数分别为 0.85 和 0.89，通过交叉校准 (RMSEcv) 计算得出的均方根误差分别为 5.25 和 1.92。所开发的方法可以快速准确地评估生物治疗产品冻干样品中的聚集情况。该研究还展示了拉曼光谱通过小瓶进行蛋白质定量的新用途。 (C) 2021 Elsevier B.V. 保留所有权利。
Quality of biotherapeutic products is of paramount importance for ensuring patient safety. Analytical tools that can facilitate rapid quality assessment of the therapeutic product at the point of care are very much in demand. In this article, we apply chemometrics based analysis of Raman spectra towards quantitative prediction of protein aggregation in lyophilized biotherapeutic products. Two commercially available therapeutic proteins, erythropoietin (EPO) and human growth hormone (HGH), have been used to demonstrate the applicability of the proposed approach. Thermally induced protein aggregation was monitored by size exclusion chromatography as well as Raman spectroscopy with a 785 nm wavelength laser. Partial least square (PLS) regression was used to analyse the Raman spectra and create a model for quantitative determination of aggregate. Satisfactory performance was observed with both EPO and HGH with R-2 of 0.91 and 0.94, cross-validation correlation coefficient of 0.85 and 0.89, and Root Mean Square Error computed from cross calibration (RMSEcv) of 5.25 and 1.92, respectively. The developed approach can enable rapid and accurate assessment of aggregation in lyophilized samples of biotherapeutic products. The study also demonstrates novel use of Raman spectroscopy for protein quantifica-tion through a vial. (C) 2021 Elsevier B.V. All rights reserved.