Raman spectroscopy provides valuable process insights for cell-derived and cellular products

Matuszczyk, Jens-Christoph; Zijlstra, Gerben; Ede, David; Ghaffari, Navid; Yuh, J.; Brivio, Veronica

doi:10.1016/j.copbio.2023.102937

Cited by 7 publications

(2 citation statements)

References 23 publications

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“…In this study, we focused on the potential of Raman spectrometry as a method for continuous analysis. Raman spectroscopy equipment generates Raman scattering light by irradiating laser light onto a sample, which carries information on the inherent oscillation frequency of molecules 9 – 11 . By detecting this light, the concentration of specific molecules can be measured.…”

Section: Introductionmentioning

confidence: 99%

Comprehensive modeling of cell culture profile using Raman spectroscopy and machine learning

Tanemura,

Kitamura,

Yamada

et al. 2023

Sci Rep

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Chinese hamster ovary (CHO) cells are widely utilized in the production of antibody drugs. To ensure the production of large quantities of antibodies that meet the required specifications, it is crucial to monitor and control the levels of metabolites comprehensively during CHO cell culture. In recent years, continuous analysis methods employing on-line/in-line techniques using Raman spectroscopy have attracted attention. While these analytical methods can nondestructively monitor culture data, constructing a highly accurate measurement model for numerous components is time-consuming, making it challenging to implement in the rapid research and development of pharmaceutical manufacturing processes. In this study, we developed a comprehensive, simple, and automated method for constructing a Raman model of various components measured by LC–MS and other techniques using machine learning with Python. Preprocessing and spectral-range optimization of data for model construction (partial least square (PLS) regression) were automated and accelerated using Bayes optimization. Subsequently, models were constructed for each component using various model construction techniques, including linear regression, ridge regression, XGBoost, and neural network. This enabled the model accuracy to be improved compared with PLS regression. This automated approach allows continuous monitoring of various parameters for over 100 components, facilitating process optimization and process monitoring of CHO cells.

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Section: Introductionmentioning

confidence: 99%

Comprehensive modeling of cell culture profile using Raman spectroscopy and machine learning

Tanemura,

Kitamura,

Yamada

et al. 2023

Sci Rep

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“…Spectroscopic methods also would be applied to strengthen the real-time IVT monitoring methods in the near future. For examples, raman spectroscopy is likely to be applied for RNA manufacturing also by measuring NTP consumption ( Matuszczyk et al, 2023 ). Circular dichroism (CD) that sensitively probes the chirality of nucleic acids would be useful to analyze nucleotide modifications and functional structure of RNA ( Valentini et al, 2022 ) during IVT and the database is currently available as a public repository ( Zhang et al, 2023 ).…”

Section: Introductionmentioning

confidence: 99%

Real-time monitoring strategies for optimization of in vitro transcription and quality control of RNA

Lee,

Song,

Kim

et al. 2023

Front. Mol. Biosci.

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RNA-based therapeutics and vaccines are opening up new avenues for modern medicine. To produce these useful RNA-based reagents, in vitro transcription (IVT) is an important reaction that primarily determines the yield and quality of the product. Therefore, IVT condition should be well optimized to achieve high yield and purity of transcribed RNAs. To this end, real-time monitoring of RNA production during IVT, which allows for fine tuning of the condition, would be required. Currently, light-up RNA aptamer and fluorescent dye pairs are considered as useful strategies to monitor IVT in real time. Fluorophore-labeled antisense probe-based methods can also be used for real-time IVT monitoring. In addition, a high-performance liquid chromatography (HPLC)-based method that can monitor IVT reagent consumption has been developed as a powerful tool to monitor IVT reaction in near real-time. This mini-review briefly introduces some strategies and examples for real-time IVT monitoring and discusses pros and cons of IVT monitoring methods.

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Use of spectroscopic process analytical technology for rapid quality evaluation during preparation of CHO cell culture media

Ou,

Cui,

Zhao

et al. 2024

Biotechnology Progress

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Media preparation parameters contribute significantly to media quality, cell culture performance, productivity, and product quality. Establishing proper media preparation procedures is critical for ensuring a robust CHO cell culture process. Process analytical technology (PAT) enables unique ways to quantify assessments and improve media quality. Here, cell culture media were prepared under a wide range of temperatures (40–80°C) and pH (7.6–10.0). Media quality profiles were compared using three real‐time PATs: Fourier‐transform infrared (FTIR) spectroscopy, Raman spectroscopy, and excitation‐emission matrix (EEM) spectroscopy. FTIR and Raman spectroscopies identified shifts in media quality under high preparation temperature (80°C) and at differing preparation pH which negatively impacted monoclonal antibody (mAb) production. In fed‐batch processes for production of three different mAbs, viable cell density (VCD) and cell viability were mostly unaffected under all media preparation temperatures, while titer and cell specific productivity of mAb decreased when cultured in basal and feed media prepared at 80°C. High feed preparation pH alone was tolerated but cell growth and productivity profiles deviated from the control condition. Further, charge variants (main, acidic, basic species) and glycosylation (G0F, afucosylation, and high mannose) were examined. Statistically significant differences were observed for one or more of these quality attributes with any shifts in media preparation. In this study, we demonstrated strong associations between media preparation conditions and cell growth, productivity, and product quality. The rapid evaluation of media by PAT implementation enabled more comprehensive understanding of different parameters on media quality and consequential effects on CHO cell culture.

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Raman spectroscopy provides valuable process insights for cell-derived and cellular products

Cited by 7 publications

References 23 publications

Comprehensive modeling of cell culture profile using Raman spectroscopy and machine learning

Comprehensive modeling of cell culture profile using Raman spectroscopy and machine learning

Real-time monitoring strategies for optimization of in vitro transcription and quality control of RNA

Use of spectroscopic process analytical technology for rapid quality evaluation during preparation of CHO cell culture media

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