by enhancing quality and
yield of lentiviral vectors
by enhancing quality and
yield of lentiviral vectors
Viruses such as lentivirus can be used to "engineer" cells either outside the body (ex vivo) or within the body (in vivo). CAR-T is a highly effective ex vivo therapy where the patients own immune cells are reprogrammed with a lentiviral vector so they can seek out and destroy cancers. With a single infusion, patients can be cured of previously intractable cancer. Lentivirus can also be used in vivo as a gene therapy to replace faulty or absent functional proteins, for example as a treatment for sickle cell disease.
Unfortunately, treatments such as CAR-T and gene therapies can be highly expensive, restricting their accessibility to patients. High end-user prices are driven by the very high production costs associated with producing components such as lentivirus. This is because the manufacturing yields for lentivirus are quite low when compared to the quantity required for the therapy.
A lot of work has been undertaken by the industry to increase lentivirus yields over the past decade. From higher efficiency transfection to increasing cell density of suspension cell lines. These advancements have made a dent in the cost of production of lentivirus, but costs still remain stubbornly high.
Current viral genome or payload plasmids use strong viral promoters such as CMV to produce the viral genome. These promoters are known to cause the expression of the payload in the production cell line, even if that payload is under the control of a tissue specific promoter. If this payload incorporates into the membrane (such as with CAR payloads) this has been shown to have a negative impact on the potency and efficacy of the therapy. Toxic or other cellular regulatory payloads are also known to have an impact on lentivirus production yield.
It is therefore desirable to minimise breakthrough expression of the lentivirus payload in the manufacturing cell line to improve quality and yield.
By making changes to the promoter which drives production of the vector plasmid, breakthrough expression of the payload plasmid is reduced by up to 100 times. By using EGFP as a reporter under a liver specific promoter in HEK293 cells, we have shown that breakthrough expression is significantly reduced. Not only the number of cells detected as positive are reduced, but also those that are positive have around a 10 fold reduction in expression.
A reduction of breakthrough expression in the manufacturing cell line will have significant, and potentially treatment enabling quality and yield benefits.
Using a novel promoter, our proprietary method for producing lentivirus can increase the quality and yield of your vector by significantly reducing breakthrough expression
The Lentitek approach is easy to integrate into your current production method.
Lentiek's approach can be applied to transient transfection as well as integrated into a stable cell line production.
Lentitek was founded in 2019 in Edinburgh, Scotland, to develop a novel approach to increase lentiviral production yields. Leveraging private funding to unlock further grant finance (Scottish Government SMART) Lentitek made rapid progress in demonstrating its technology through a successful proof of concept program. This early data coupled with the clear potential was recognised by the company winning the prestigious Scottish EDGE industrial biotechnology innovation top prize which included a significant cash injection. Results from the PoC combined with continuous development of the platform through 2022-23 have driven further private investment and the company is now actively seeking application partners to exploit the technology.
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