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Article

Dielectric Spectroscopy to Improve the Production of rAAV Used in Gene Therapy

1
iBET—Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901 Oeiras, Portugal
2
ITQB-NOVA, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal
3
Voyager Therapeutics, 75 Sidney St, Cambridge, MA 02139, USA
*
Author to whom correspondence should be addressed.
Processes 2020, 8(11), 1456; https://doi.org/10.3390/pr8111456
Received: 2 October 2020 / Revised: 7 November 2020 / Accepted: 10 November 2020 / Published: 13 November 2020
(This article belongs to the Special Issue Measurement Technologies for up- and Downstream Bioprocessing)
The insect cell-baculovirus expression vector system is an established method for large scale recombinant adeno-associated virus (rAAV) production, largely due to its scalability and high volumetric productivities. During rAAV production it is critical to monitor process parameters such as Spodoptera frugiperda (Sf9) cell concentration, infection timing, and cell harvest viabilities since they can have a significant influence on rAAV productivity and product quality. Herein we developed the use of dielectric spectroscopy as a process analytical technology (PAT) tool used to continuously monitor the production of rAAV in 2 L stirred tank bioreactors, achieving enhanced control over the production process. This study resulted in improved manufacturing robustness through continuous monitoring of cell culture parameters, eliminating sampling needs, increasing the accuracy of infection timing, and reliably estimating the time of harvest. To increase the accuracy of baculovirus infection timing, the cell growth/permittivity model was coupled to a feedback loop with real-time monitoring. This system was able to predict baculovirus infection timing up to 24 h in advance for greatly improved accuracy of infection and ensuring consistent high rAAV productivities. Furthermore, predictive models were developed based on the dielectric measurements of the culture. These multiple linear regression-based models resulted in correlation coefficients (Q2) of 0.89 for viable cell concentration, 0.97 for viability, and 0.92 for cell diameter. Finally, models were developed to predict rAAV titer providing the capability to distinguish in real time between high and low titer production batches. View Full-Text
Keywords: AAV—adeno-associated virus; insect cell-baculovirus; gene therapy; cell culture monitoring; process analytical technology; dielectric spectroscopy AAV—adeno-associated virus; insect cell-baculovirus; gene therapy; cell culture monitoring; process analytical technology; dielectric spectroscopy
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MDPI and ACS Style

Pais, D.A.M.; Brown, C.; Neuman, A.; Mathur, K.; Isidro, I.A.; Alves, P.M.; Slade, P.G. Dielectric Spectroscopy to Improve the Production of rAAV Used in Gene Therapy. Processes 2020, 8, 1456. https://doi.org/10.3390/pr8111456

AMA Style

Pais DAM, Brown C, Neuman A, Mathur K, Isidro IA, Alves PM, Slade PG. Dielectric Spectroscopy to Improve the Production of rAAV Used in Gene Therapy. Processes. 2020; 8(11):1456. https://doi.org/10.3390/pr8111456

Chicago/Turabian Style

Pais, Daniel A.M., Chris Brown, Anastasia Neuman, Krishanu Mathur, Inês A. Isidro, Paula M. Alves, and Peter G. Slade 2020. "Dielectric Spectroscopy to Improve the Production of rAAV Used in Gene Therapy" Processes 8, no. 11: 1456. https://doi.org/10.3390/pr8111456

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