EVs share similar sizes, biophysical characteristics and even biogenesis pathways with enveloped viruses, which are currently used as key ingredients in a number of viral vectors and licensed vaccine products. All these expression systems, like most cells and cellular tissues, are known to spontaneously release extracellular vesicles (EVs). Viral vectors and vaccines are produced in different platforms, from traditional embryonated chicken eggs to more advanced cell cultures. Many infectious diseases are controlled using vaccines designed from subunits or whole viral structures, whereas other genetic diseases and cancers are being treated by viruses used as vehicles for delivering genetic material in gene therapy or as therapeutic agents in virotherapy protocols.
Finally, the process economy impact of Capto Core 700 compared with Sepharose 4 Fast Flow, a GF medium often used in the vaccine manufacturing industry today, was also investigated.Viral vectors and viral vaccines are invaluable tools in prevention and treatment of diseases. The approach taken for designing a purification process for flu vaccine is also described (Fig 1). An experiment to determine the binding capacity and window of operation with Capto Core 700 for Madin- Darby canine kidney (MDCK) cell protein was performed using PreDictor 96-well plates for high-throughput process development. The aim of this work was to evaluate Capto Core 700 chromatography medium as an alternative to current chromatography technologies used in vaccine processes. Using Capto Core 700 allows higher flow rates (1) and significantly higher sample loads than traditional GF (typically several column volumes compared with 0.1 to 0.3 column volumes in GF). These internalized ligands bind various contaminants strongly over a wide range of pH and salt concentrations. The octylamine ligands in the core of the bead are multimodal, being both hydrophobic and positively charged. Viruses and other large entities with a molecular mass (M r ) greater than approximately 700 000 (700 kDa) are excluded and are collected in the chromatography flowthrough. Small contaminant molecules enter into the beads where they are captured. The Capto Core 700 medium has a core bead design and consists of an inactive shell and a ligand-activated core.
Three alternative process streams for the purification of influenza A/H1N1 were evaluated in this study. In GF, the low productivity relates to low flow rates and limited sample loads. In these processes, both sucrose density gradient ultracentrifugation and GF have limitations in, for example, scalability and productivity. The methods for purification processes have typically involved a combination of sucrose density gradient ultracentrifugation, ultrafiltration/diafiltration (UF/DF) with hollow-fiber membranes, and chromatography using affinity-, ion exchange-, and/or gel filtration (GF) media. However, to meet the needs for pandemic preparedness and scalability of vaccine productions, cell-based processes are being developed and implemented to a greater extent in the industry. Introduction Influenza vaccine has historically and is today primarily produced in embryonated chicken eggs. This study shows that while offering the same purity as gel filtration, Capto Core 700 enables significant improvements in productivity and process economy.
Here, the performance of Capto Core 700 in three different processes for the purification of influenza H1N1 virus from infected mammalian cells was evaluated and compared. The size separating mode of operation could be compared to gel filtration (size exclusion chromatography), which is a common approach for polishing steps in vaccine processes. Capto Core 700 is built on the core bead technology which allows for dual functionality combining size separation with binding chromatography. The medium is designed to be used in flowthrough mode for large targets (> M r 700 000) while scavenging smaller contaminants. GE Healthcare Life Sciences Application note 29-0003-34 AA Vaccines Purification of influenza A/H1N1 using Capto Core 700 Capto Core 700 is a chromatography medium (resin) optimized for purification of viruses and other large biomolecules.
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