Brunner Matthias, Brosig Philipp, Losing Monika, Kunzelmann Marco, Calvet Amandine, Stiefel Fabian, Bechmann Jan, Unsoeld Andreas, Schaub Jochen
Bioprocess Development Biologicals Boehringer Ingelheim Pharma GmbH & Co. KG Biberach Germany.
Analytical Development Biologicals Boehringer Ingelheim Pharma GmbH & Co. KG Biberach Germany.
Eng Life Sci. 2019 Aug 22;19(10):666-680. doi: 10.1002/elsc.201900050. eCollection 2019 Oct.
Biopharmaceutical manufacturing processes can be affected by variability in cell culture media, e.g. caused by raw material impurities. Although efforts have been made in industry and academia to characterize cell culture media and raw materials with advanced analytics, the process of industrial cell culture media preparation itself has not been reported so far. Within this publication, we first compare mid-infrared and two-dimensional fluorescence spectroscopy with respect to their suitability as online monitoring tools during cell culture media preparation, followed by a thorough assessment of the impact of preparation parameters on media quality. Through the application of spectroscopic methods, we can show that media variability and its corresponding root cause can be detected online during the preparation process. This methodology is a powerful tool to avoid batch failure and is a valuable technology for media troubleshooting activities. Moreover, in a design of experiments approach, including additional liquid chromatography-mass spectrometry analytics, it is shown that variable preparation parameters such as temperature, power input and preparation time can have a strong impact on the physico-chemical composition of the media. The effect on cell culture process performance and product quality in subsequent fed-batch processes was also investigated. The presented results reveal the need for online spectroscopic methods during the preparation process and show that media variability can already be introduced by variation in media preparation parameters, with a potential impact on scale-up to a commercial manufacturing process.
生物制药生产过程可能会受到细胞培养基变异性的影响，例如由原材料杂质引起的变异性。尽管工业界和学术界已努力使用先进的分析方法来表征细胞培养基和原材料，但迄今为止，工业细胞培养基制备过程本身尚未见报道。在本出版物中，我们首先比较了中红外光谱和二维荧光光谱在细胞培养基制备过程中作为在线监测工具的适用性，随后全面评估了制备参数对培养基质量的影响。通过应用光谱方法，我们可以证明在制备过程中可以在线检测到培养基的变异性及其相应的根本原因。这种方法是避免批次失败的有力工具，也是用于培养基故障排除活动的宝贵技术。此外，在包括额外的液相色谱 - 质谱分析的实验设计方法中，结果表明诸如温度、功率输入和制备时间等可变制备参数会对培养基的物理化学组成产生强烈影响。还研究了其对后续补料分批培养过程中细胞培养过程性能和产品质量的影响。所呈现的结果揭示了在制备过程中需要在线光谱方法，并表明培养基变异性可能已经由培养基制备参数的变化引入，这对扩大到商业制造过程具有潜在影响。
