Proton-transfer-reaction mass spectrometry (PTR-MS) for online monitoring of glucose depletion and cell concentrations in HEK 293 gene therapy processes

Proton-transfer-reaction mass spectrometry (PTR-MS) for online monitoring of glucose depletion and cell concentrations in HEK 293 gene therapy processes

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 pro...

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Bibliographic Details
Published in:Biotechnology letters Vol. 44; no. 1; pp. 77 - 88
Main Authors: Bayer, Benjamin, Maccani, Andreas, Jahn, Johanna, Duerkop, Mark, Kapeller, Ewald, Pletzenauer, Robert, Kraus, Barbara, Striedner, Gerald, Hernandez Bort, Juan A.
Format: Journal Article
Language:English
Published: Dordrecht Springer Netherlands 01-01-2022
Springer Nature B.V
Subjects:
Acetaldehyde
Applied Microbiology
Biochemistry
Biomedical and Life Sciences
Biotechnology
Capacitance
Cell culture
Depletion
Evaluation
Gene therapy
Genetic Therapy
Glucose
HEK293 Cells
Humans
Information processing
Life Sciences
Mass spectrometry
Mass Spectrometry - methods
Mass spectroscopy
Microbiology
Monitoring
Organic compounds
Original Research Paper
Process control
Process controls
Protons
Reproducibility of Results
Scientific imaging
Spectroscopy
Transfection
VOCs
Volatile organic compounds
Volatile Organic Compounds - analysis
Workflow
PTR-MS monitoring
HEK 293 cell culture monitoring
rAAV process
Glucose online monitoring
Cell culture PAT
Biomass soft sensor
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Summary: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.
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ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-021-03205-y