Real‐Time VCC Monitoring and Forecasting in HEK‐Cell‐Based rAAV Vector Production Using Capacitance Spectroscopy
ABSTRACT Recombinant adeno‐associated virus (rAAV) vector production is a complex process in which the robust cultivation of human embryonic kidney cells (HEK293) plays a critical role in generating high‐quality viral vectors. Tracking the viable cell concentration (VCC) during upstream production i...
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| Format: | Article |
| Language: | English |
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Wiley-VCH
2025-02-01
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| Series: | Engineering in Life Sciences |
| Online Access: | https://doi.org/10.1002/elsc.70004 |
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| author | Rafael Machleid Suneetha Nunna Ajith George Jonas Austerjost Magda Tomala Izabella Surowiec |
| author_facet | Rafael Machleid Suneetha Nunna Ajith George Jonas Austerjost Magda Tomala Izabella Surowiec |
| author_sort | Rafael Machleid |
| collection | DOAJ |
| description | ABSTRACT Recombinant adeno‐associated virus (rAAV) vector production is a complex process in which the robust cultivation of human embryonic kidney cells (HEK293) plays a critical role in generating high‐quality viral vectors. Tracking the viable cell concentration (VCC) during upstream production is essential for process monitoring and for implementing actions that ensure optimal process management. The advent of inline capacitance probes has introduced a crucial process analytical technology (PAT) tool for real‐time VCC measurement. Here, we present the development and application of a method for real‐time monitoring of VCC in HEK293‐based rAAV vector production. In a first step, BioPAT Viamass probes were used to record capacitance data of individual 10 L rAAV‐8 batches within a frequency range of 50 kHz–20 MHz. Based on the capacitance data, a linear single‐frequency model and an orthogonal partial least square (OPLS) multifrequency model for VCC prediction were developed. Subsequently, these models were deployed inline, and predictions were exposed into BioPAT MFCS bioprocess control software, enabling real‐time VCC monitoring in subsequent rAAV‐8 production batches. In addition, the continuous VCC signal was used as input for an exponential cell growth model that was deployed inline to provide accurate real‐time forecasting of the transfection time point. To the best of our knowledge, this is the first example of inline deployment of VCC and Time‐Till‐Transfection predictive models to the bioprocess control system for real‐time monitoring and forecasting of these parameters in HEK‐cell‐based transient rAAV vector production. |
| format | Article |
| id | doaj-art-5ef70536d4a54256b9da277bcb08cc30 |
| institution | DOAJ |
| issn | 1618-0240 1618-2863 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Wiley-VCH |
| record_format | Article |
| series | Engineering in Life Sciences |
| spelling | doaj-art-5ef70536d4a54256b9da277bcb08cc302025-08-20T02:54:43ZengWiley-VCHEngineering in Life Sciences1618-02401618-28632025-02-01252n/an/a10.1002/elsc.70004Real‐Time VCC Monitoring and Forecasting in HEK‐Cell‐Based rAAV Vector Production Using Capacitance SpectroscopyRafael Machleid0Suneetha Nunna1Ajith George2Jonas Austerjost3Magda Tomala4Izabella Surowiec5Sartorius Stedim Biotech GmbH Advanced Analytics & Spectroscopy, Embedded Hardware Göttingen GermanySartorius Stedim Biotech GmbH Application Testing Separation Consumables – Upstream Göttingen GermanySartorius Stedim Biotech GmbH Advanced Analytics & Spectroscopy, Embedded Hardware Göttingen GermanySartorius Stedim Biotech Advanced BioAnalytics, Corporate Research Göttingen GermanySartorius Stedim Biotech GmbH Application Testing Separation Consumables – Upstream Göttingen GermanySartorius Stedim Data Analytics AB Advanced Analytics & Spectroscopy, Embedded Hardware Umeå SwedenABSTRACT Recombinant adeno‐associated virus (rAAV) vector production is a complex process in which the robust cultivation of human embryonic kidney cells (HEK293) plays a critical role in generating high‐quality viral vectors. Tracking the viable cell concentration (VCC) during upstream production is essential for process monitoring and for implementing actions that ensure optimal process management. The advent of inline capacitance probes has introduced a crucial process analytical technology (PAT) tool for real‐time VCC measurement. Here, we present the development and application of a method for real‐time monitoring of VCC in HEK293‐based rAAV vector production. In a first step, BioPAT Viamass probes were used to record capacitance data of individual 10 L rAAV‐8 batches within a frequency range of 50 kHz–20 MHz. Based on the capacitance data, a linear single‐frequency model and an orthogonal partial least square (OPLS) multifrequency model for VCC prediction were developed. Subsequently, these models were deployed inline, and predictions were exposed into BioPAT MFCS bioprocess control software, enabling real‐time VCC monitoring in subsequent rAAV‐8 production batches. In addition, the continuous VCC signal was used as input for an exponential cell growth model that was deployed inline to provide accurate real‐time forecasting of the transfection time point. To the best of our knowledge, this is the first example of inline deployment of VCC and Time‐Till‐Transfection predictive models to the bioprocess control system for real‐time monitoring and forecasting of these parameters in HEK‐cell‐based transient rAAV vector production.https://doi.org/10.1002/elsc.70004 |
| spellingShingle | Rafael Machleid Suneetha Nunna Ajith George Jonas Austerjost Magda Tomala Izabella Surowiec Real‐Time VCC Monitoring and Forecasting in HEK‐Cell‐Based rAAV Vector Production Using Capacitance Spectroscopy Engineering in Life Sciences |
| title | Real‐Time VCC Monitoring and Forecasting in HEK‐Cell‐Based rAAV Vector Production Using Capacitance Spectroscopy |
| title_full | Real‐Time VCC Monitoring and Forecasting in HEK‐Cell‐Based rAAV Vector Production Using Capacitance Spectroscopy |
| title_fullStr | Real‐Time VCC Monitoring and Forecasting in HEK‐Cell‐Based rAAV Vector Production Using Capacitance Spectroscopy |
| title_full_unstemmed | Real‐Time VCC Monitoring and Forecasting in HEK‐Cell‐Based rAAV Vector Production Using Capacitance Spectroscopy |
| title_short | Real‐Time VCC Monitoring and Forecasting in HEK‐Cell‐Based rAAV Vector Production Using Capacitance Spectroscopy |
| title_sort | real time vcc monitoring and forecasting in hek cell based raav vector production using capacitance spectroscopy |
| url | https://doi.org/10.1002/elsc.70004 |
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