Characterization and Qualification of Viral, bacterial and Plasmid DNA Vaccines
Many manufacturers face the challenge of testing virus stocks in cell-based assay systems. MDS can design accurate, efficient testing strategies in concert with your manufacturing experts. If neutralizing antibodies are needed, we can work to qualify the assay systems, ensuring reliable results.
Assays for Product Manufacture of Plasmid DNA Vaccines to ensure that both the MCBs and WCBs are:
- Free from bacteriophage contamination
- Free from adventitious agent contamination
- Genetically stable stability of the plasmid DNA in the WCB.
Since processing cell lines and/or virus seed stocks may at some point have been exposed to animal-derived materials,
MDS additionally offers testing for bovine and porcine viruses. These assays are designed per instructions in the U.S. Code of Federal Regulations, Animals and Animal Products, Title 9 (9 CFR).
- Visual appearance
- Plasmid concentration.
- The fraction of plasmid in supercoiled conformation
- Endotoxin (LAL)
- The presence of bacterial host cell macromolecules including
- Aagarose gel electrophoresis of plasmid DNA after restriction enzyme digestion
- in vitro measures of transfection efficiency
- in vivo assays of DNA vaccine immunogenicity.
- DNA sequence of the insert gene
- General safety
- Endotoxin (LAL)
- Changes to the Insert or Vector
- DNA Sequence Analysis
- Toxicity and Biodistribution
- Antigen-specific antibody titers
- Seroconversion rates
- Activation of cytokine secreting cell
- Cell-mediated immune responses
- Autoimmunity: DNA vaccination can activate autoreactive B cells to secrete IgG anti-DNA autoantibodies.
- Q-PCR for Integration, Biodistribution, Persistence, and Integration Analysis Specifically designed PCR primers may be used to confirm integration and identify genomic integration sites.
- Tumorigenicity if insertion reduces the activity of a tumor suppressor or increases the activity of an oncogene.
- Local Reactogenicity and Systemic Toxicity Studies