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Monitoring lentiviral vector production kinetics using online permittivity measurements

Paper ID Volume ID Publish Year Pages File Format Full-Text
3827 191 2011 10 PDF Available
Title
Monitoring lentiviral vector production kinetics using online permittivity measurements
Abstract

Lentiviral vectors (LVs) are effective delivery vehicles that have been successfully used in gene and cell therapy. LVs are most commonly produced via the transient transfection of several plasmid constructs in adherent cell cultures. Recently, we described an efficient and scalable LV production in serum-free suspension cultures. To further facilitate the translation of LV-based interventions to the clinic, the robustness of the production process needs to be ensured to ultimately achieve a specified quality and quantity of LV production lots. However, routine processes are largely empirical, and strategies to monitor LV production kinetics in real-time have not yet been described.In this work, in situ real-time permittivity measurements were assessed to document the production of LVs. Characteristic process phases that were closely associated with LV production kinetics were identified. The permittivity signal evolution was interpreted by exploiting various independent online and offline monitoring measurements. Cellular membrane properties and, to a lesser extent, cell size were the main factors contributing to the permittivity variations. It is concluded that the permittivity-related parameters can be used for the detection of viral release, allowing real-time assessment of process performance. The technology should thus greatly facilitate process development and optimization.

Research highlights► In situ real-time permittivity measurements were assessed to document the production of lentiviral vectors (LVs). ► Characteristic process phases associated with LV production kinetics were identified. ► Cellular membrane properties and cell size caused the permittivity variations. ► The release of LVs in the culture supernatant can be monitored in real time by permittivity measurements.

Keywords
Lentiviral vector production; HEK293; Perfusion culture; Online monitoring; Permittivity measurements; Dielectric spectroscopy; Process Analytical Technology (PAT)
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Monitoring lentiviral vector production kinetics using online permittivity measurements
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 54, Issue 1, 15 March 2011, Pages 16–25
Authors
, , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us