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.
Biopharmaceutical Applications Guide - Vaccines
Brochure: Vaccine Bioprocessing Handbook
Mini-Handbook: Viral Vector Vaccine Bioprocessing
Webinar: Process Development Guidance for AAV and Lentivirus Manufacturing Based on Cost Modeling
Webinar: Sf-RVN Cell Line & CD Media Platform, A Production Platform for VLP and AAV
White Paper: Developing an Accelerated and More Cost-Effective Single-Use Adenoviral Vector Vaccine Manufacturing Process
Technical Article: Purification or Removal of Viruses including Adeno-associated Virus
Article: Robust Harvest Clarification For Adeno-Associated Viral Vectors Via Depth Filtration
Article: Clarification of vaccines: An overview of filter based technology trends and best practices
Article: Filter-Based Clarification of Viral Vaccines and Vectors Biosafety
Article: How to Ensure Biosafety Of Novel Viral Therapies
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.
Ensure robust scalability.
Achieve yield, efficiency and pDNA recovery goals while ensuring robust impurity removal
We offer the industry’s highest quality sterile filtered liquid capabilities, supplying ready-to-use cell culture media, buffers, CIP and SIP products from GMP facilities worldwide to optimize your biopharma production.
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