| Title: | Proton-transfer-reaction mass spectrometry (PTR-MS) for online monitoring of glucose depletion and cell concentrations in HEK 293 gene therapy processes |
| Author(s): | Bayer B; Maccani A; Jahn J; Duerkop M; Kapeller E; Pletzenauer R; Kraus B; Striedner G; Hernandez Bort JA; |
| Address: | "Novasign GmbH, Vienna, Austria. University of Natural Resources and Life Sciences, Vienna, Austria. Gene Therapy Process Development, Baxalta Innovations GmbH, a Part of Takeda Companies, Uferstrasse 15, 2304, Orth an der Donau, Austria. Gene Therapy Process Development, Baxalta Innovations GmbH, a Part of Takeda Companies, Uferstrasse 15, 2304, Orth an der Donau, Austria. juan.hernandezbort@takeda.com" |
| DOI: | 10.1007/s10529-021-03205-y |
| ISSN/ISBN: | 1573-6776 (Electronic) 0141-5492 (Print) 0141-5492 (Linking) |
| Abstract: | "OBJECTIVES: The applicability of proton-transfer-reaction mass spectrometry (PTR-MS) as a versatile online monitoring tool to increase consistency and robustness for recombinant adeno-associated virus (rAAV) producing HEK 293 bioprocesses was evaluated. We present a structured workflow to extract process relevant information from PTR-MS data. RESULTS: Reproducibility of volatile organic compound (VOC) measurements was demonstrated with spiking experiments and the process data sets used for applicability evaluation consisted of HEK 293 cell culture triplicates with and without transfection. The developed data workflow enabled the identification of six VOCs, of which two were used to develop a soft sensor providing better real-time estimates than the conventional capacitance sensor. Acetaldehyde, another VOC, provides online process information about glucose depletion that can directly be used for process control purposes. CONCLUSIONS: The potential of PTR-MS for HEK 293 cell culture monitoring has been shown. VOC data derived information can be used to develop soft sensors and to directly set up new process control strategies" |
| Keywords: | Genetic Therapy Glucose HEK293 Cells Humans Mass Spectrometry/methods *Protons Reproducibility of Results *Volatile Organic Compounds/analysis Biomass soft sensor Cell culture PAT Glucose online monitoring HEK 293 cell culture monitoring PTR-MS monitoring; |
| Notes: | "MedlineBayer, Benjamin Maccani, Andreas Jahn, Johanna Duerkop, Mark Kapeller, Ewald Pletzenauer, Robert Kraus, Barbara Striedner, Gerald Hernandez Bort, Juan A eng Netherlands 2021/11/13 Biotechnol Lett. 2022 Jan; 44(1):77-88. doi: 10.1007/s10529-021-03205-y. Epub 2021 Nov 12" |
