Supporting Adenoviral Vector Development for Gene Therapy and Vaccine Research Adenoviral vectors offer high levels of transgene expression in broad cell types, including both dividing and non-dividing cells. They are the method of choice for applications requiring fast and transient gene expression for large genes or multiple genes, with a capacity of up to 7.5 kb. Additionally, adenoviral vectors are widely used in vaccine development due to their ability to induce a strong immune response.
Revvity’s gene delivery key advantages
- Expert Consultancy: Personalized planning and consultation to help meet the unique needs of each project.
- Quality and Consistency: Standards for reliable batch-to-batch quality.
- Customizable QC Packages: Tailored to work with the specific requirements of your research or clinical application.
- Scalability: From research-grade to large-scale production.
For research use only. Not for use in diagnostic procedures.
General Adenovirus Characteristics
- Transient Gene Expression: Effective in vivo and in vitro.
- No Host Genome Integration: Remains epichromosomal, avoiding interference with host genes.
- Large Packaging Capacity: Up to 7.5 kb, allowing for multiple gene expression cassettes.
- High Efficiency: Reliable gene delivery into both dividing and non-dividing cells.
- Fast Results: Gene expression within 24 hours.
Adenoviral Vector Engineering
Adenoviruses are non-toxic, non-integrating, non-enveloped viruses with linear double-stranded DNA. Our adenoviral vectors are based on the Ad5 serotype, which uses the Coxsackie-Adenovirus Receptor (CAR) to enter cells. To increase packaging capacity, the E1 and E3 genes are deleted, preventing viral replication and making space for your gene of interest.
Vector Construction:
To construct an adenoviral vector, the gene of interest (GOI) is cloned into an adenoviral shuttle plasmid. This plasmid undergoes FLP-mediated recombination with a BAC vector in BA5-FRT cells, resulting in a linear Ad5-GOI construct.
Vector Production:
The linear construct is transfected into HEK293 or CAP producer cells to generate a low-titer virus stock. This stock is then amplified, concentrated, and purified for QC testing, and finally filled and finished for delivery.
Adenoviral Vector Manufacturing Process
Day 1: Transfection of HEK293 cells with the linear DNA containing Ad5-GOI.
Days 2-3: Incubation and collection of virus from supernatant and cell lysates. Subsequent Rounds: Amplification of the virus to the required scale, followed by purification using CsCl or AdenoOne purification kits.
QC and Analytics
We offer various quality control (QC) packages to assess the characteristics of your adenoviral vectors. Standard and optional QC methods are available, including genome titration, capsid titration, purity checks, and more. Additional assays can be customized for your specific needs.
Scales and Timelines
Revvity offers multiple production stages to accommodate different project requirements, from initial cloning and vector development to large-scale production and simple amplification of existing vector stocks. Our production timelines and scales are adaptable to help meet the specific needs of your project.
