I1 - Vector Product Engineering, Development and Manufacturing (excluding AAV)
1856: A Novel, Synthetic DNA Alternative for Lentiviral Vectors Manufacturing
Type: Poster Session
Poster Board Number: 1856
Presentation Details
Session Title: Friday Posters: Vector Product Engineering, Development, and Manufacturing
Lentivirus vectors (LVV) represent vital products and tools within the cell and gene therapy domains. LVV are conventionally produced in cellular expression systems upon transfection with plasmids of bacterial origin that encode the gene-of-interest (GOI), packaging proteins, and envelope proteins. Scaling up the production of these plasmids is costly, and time-consuming.
We have developed a novel, optimized synthetic linear double-stranded DNA with customizable hairpin-ended structures for LVV production. Our cell-free, enzymatic production process is readily scalable and results in high-fidelity, high-yield, and high-purity DNA in a short timeframe. Additionally, our synthetic DNA constructs are devoid of bacterial backbone, thereby reducing the risk of introducing non-relevant and antibiotic resistance elements into the LVV production process - a significant safety and regulatory concern. Our DNA platform encodes the elements of a 3rd generation lentivirus system. LVV were produced by transient cell transfection at optimal conditions determined by Design of Experiments. Functionality and titer of the produced LVV was evaluated by cell transduction assay and p24 ELISA.
Here, we show that synthetic DNA constructs codifying for the GOI, Rev, Gag-Pol, and VDV-G generated through our enzymatic, cell-free process are produced at high purity. GOIs encoded in our DNA platform include reporters of different size, anti-CD19 Chimeric Antigen Receptor (CAR), under the control of different promoters. All LVV produced were functional, effectively transducing HEK293T cells. CD19 CAR was also effectively expressed at the surface of Jurkat cells upon transduction with the produced LVV, further demonstrating the feasibility of our DNA in applications for CAR-T cell therapies development. In conclusion, our synthetic DNA platform can be used as an alternative to bacterial plasmids in LVV production.
Plain Language Summary
As more gene therapies are approved and others enter clinical trials, lentiviral vector (LLV) manufacturing faces challenges to scale to meet demand. We have developed a novel, optimized synthetic linear double-stranded DNA with customizable hairpin-ended structures for LVV production. Our cell-free, enzymatic production process is readily scalable and results in high-fidelity, high-yield, and high-purity DNA in a short timeframe. Our synthetic DNA constructs are devoid of bacterial backbone, thereby reducing the risk of introducing non-relevant and antibiotic-resistant elements into the LVV production process. We show that our novel DNA is an innovative and robust solution to help scale LLV manufacturing.
Andreia M. Silva, Fabian Trick, David Wilson, Ivana Pastierikova, Luca Distefano, Marco Guarrera, Martin Cusack, Alexander Pekarsky, Jorge Omar Yanez Cuna, Joel de Beer
Anjarium Biosciences AG, Schlieren, Switzerland"
Find This Session