Fig 1. Process Train for Viral Vector Vaccines
A live vector vaccine uses an attenuated or harmless microorganism such as an adenovirus to transport portions of an antigen to stimulate an immune response. Vectored vaccines are capable of inducing potent cell-mediated immunity, which is essential for complex disease like AIDS, malaria, and cancer among others.
While the manufacturing process for vaccine vectors is fairly templated, some challenges may arise, since several different viruses with varying properties can be used. For large viral vectors, process sterility is critical due to yield loss associated with sterile filtration. There are also challenges with vector aggregation and stability.
In addition, many Phase I/II processes involve adherent cell cultures; as a result, scaling to Phase III and commercial manufacturing can require process adjustments to achieve production targets. In later phases, product yield and purity are critical, as high dosage titers are required in the final product.
Upstream culture processes developed for manufacturing of viral vector vaccines must be optimized to meet productivity requirements. This optimization includes the cell lysis and clarification steps which are essential for removal of cells and cell debris and to ensure a robust vector harvest. The upstream process is only successful, however, if it can be reliably scaled in order to meet anticipated market demand.
Nucleic acids from lysed cells are a common contaminant in viral vector vaccine processes. Regulations require that the level of carry-over host cell nucleic acid be below 10 ng/dose of attenuated viral vaccine. Benzonase® endonuclease treatment followed by tangential flow filtration is a robust and powerful combination to degrade and remove residual nucleic acid components.
Small-scale clinical lots are typically purified using CsCl-based density gradient ultracentrifugation, while large-scale production requires a two- or three-step chromatography process. Anion exchange is typically used to remove HCP, DNA, RNA, and other major contaminants, while size exclusion chromatography is used for trace contaminant removal.
Sterile filtration ensures the sterility of the final formulated product and patient safety. A filter pore size of 0.22 µm or less is required to eliminate microbial contaminants. A key consideration for the sterile filtration process is the level of viral aggregates. These aggregates need to be controlled by optimizing the formulation, otherwise, the sterile filtration process will be challenging with the potential for high losses in yield.
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The manufacturing process for adenovirus vectors is straightforward and fairly templated; a general outline is explained in this handbook.
Our collaboration with the Jenner Institute has resulted in a single-use, scalable GMP template for manufacture of adenoviral-based vaccines. Learn more on this faster and more cost-effective approach to vaccine manufacturing.
This Application Note summarizes the benefits of incorporating Benzonase® endonuclease in a polio virus type 3 production process.
We demonstrate that depth filters clarify AAV vectors, helping to overcome the unique separation challenges presented by these important vectors for gene therapy.
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