Automized inline monitoring in perfused mammalian cell culture by MIR spectroscopy without calibration model building

IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Hannah Marienberg, Nicole Desch, Vitalii Mozin, Lorenz Sykora-Mirle, Anja Müller, Andreas Roth, Mathias Käfer, Rüdiger Neef
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Abstract

Process Analytical Technologies (PATs) are taking a key role in the run for automatization in the biopharmaceutical industry. Spectroscopic methods such as Raman spectroscopy or mid-infrared (MIR) spectroscopy are getting more recognition in the recent years for inline monitoring of bioprocesses due to their ability to measure various molecules simultaneously. However, their dependency on laborious model calibration making them a challenge to implement. In this study, a novel one-point calibration that requires a single reference point prior to the inline monitoring of glucose and lactate in bioprocesses with MIR spectroscopy is assessed with 22 mammalian cell perfusion (PER) processes in two different scales and four different products. Concentrations are predicted over all PERs runs with a root mean square error (RMSE) of 0.29 g/L for glucose and 0.24 g/L for lactate, respectively. For comparison conventional partial least square regression (PLSR) models were used and trained with spectroscopic data from six bioreactor runs in two different scales and three products. The general accuracy of those models (RMSE of 0.41 g/L for glucose and 0.16 g/L for lactate) are in the range of the accuracy of the one-point calibration. This shows the potential of the one-point calibration as an approach making spectroscopy more accessible for bioprocess development.

Abstract Image

利用近红外光谱对灌注哺乳动物细胞培养进行自动在线监测,无需建立校准模型
过程分析技术(PATs)在生物制药行业的自动化进程中发挥着关键作用。近年来,拉曼光谱或中红外光谱等光谱方法由于能够同时测量各种分子而在生物过程的在线监测中得到越来越多的认可。然而,它们对费力的模型校准的依赖使它们成为实现的挑战。在这项研究中,在使用MIR光谱在线监测生物过程中的葡萄糖和乳酸之前,需要一个单一参考点的一种新颖的单点校准,在两种不同的尺度和四种不同的产品中对22种哺乳动物细胞灌注(PER)过程进行了评估。在所有的per运行中,浓度预测的均方根误差(RMSE)分别为葡萄糖0.29 g/L和乳酸0.24 g/L。为了进行比较,使用了传统的偏最小二乘回归(PLSR)模型,并对六个生物反应器在两种不同规模和三种产品上运行的光谱数据进行了训练。这些模型的一般精度(葡萄糖的RMSE为0.41 g/L,乳酸的RMSE为0.16 g/L)在一点校准的精度范围内。这显示了一点校准作为一种使光谱学更容易用于生物过程开发的方法的潜力。
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来源期刊
Engineering in Life Sciences
Engineering in Life Sciences 工程技术-生物工程与应用微生物
CiteScore
6.40
自引率
3.70%
发文量
81
审稿时长
3 months
期刊介绍: Engineering in Life Sciences (ELS) focuses on engineering principles and innovations in life sciences and biotechnology. Life sciences and biotechnology covered in ELS encompass the use of biomolecules (e.g. proteins/enzymes), cells (microbial, plant and mammalian origins) and biomaterials for biosynthesis, biotransformation, cell-based treatment and bio-based solutions in industrial and pharmaceutical biotechnologies as well as in biomedicine. ELS especially aims to promote interdisciplinary collaborations among biologists, biotechnologists and engineers for quantitative understanding and holistic engineering (design-built-test) of biological parts and processes in the different application areas.
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