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05 July 2022

Developing Stable Lentiviral Cell Lines

Editorial /International Biopharmaceutical Industry/05 July 2022/Issue Volume 5, Issue 2

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Corinne Branciaroli & Dr. Katie Roberts


Lentiviral vectors (LVV) are commonly used as gene delivery tools for cell and gene therapies, notably chimeric antigen receptor (CAR) T cell therapies. Like other retroviruses, lentiviruses can convert their single-stranded RNA genome into double-stranded DNA when integrating into the genome of a cell. Unlike other retroviruses, lentiviruses can transduce non-dividing and quiescent cells, which makes them highly suitable for use in cell therapy.

The positive effects of novel cell therapies on malignancies, followed by the approvals granted by regulatory bodies such as the FDA and EMA, led to an increased interest in cell therapies. This in turn has led to an increased number of clinical trials and a growing demand for lentiviral manufacture.

Process scalability and robustness are therefore essential to ensure consistent and reproducible production, maximise yields and lower costs. Lentiviral vectors are often produced in HEK293 cells transiently transfected with four (or more) plasmids that contain all the genetic material required for vector production (GagPol, VSV-G, Rev and Lentiviral genome gene of interest (LV_Genome_GOI)).

However, while transient systems are useful in some contexts, transient systems can often be subject to more variability, a higher cost and larger amounts of complexity than stable systems. The development of stable packaging and producer cell lines, which integrate some or all of the lentivirus elements into the host cell genome, could provide a simpler, more scalable alternative to transient systems, reducing the variability associated with transient transfection and generating more consistent titres across manufacturing runs.

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