Downstream processing remains one of the main bottlenecks in Adenoviral vector manufacturing. At BIA Separations, a Sartorius company, we offer a platform for purification of Adenoviral vaccines using market leading CIM® monolithic chromatographic columns and an analytical toolbox for process monitoring in Adenoviral vaccine production.

Simplified purification of Adenovirus consists of typical downstream steps, including combined lysis, clarification, TFF and a chromatographic capture on pre-packed monolithic CIMmultus QA column.

TFF Retentate is prepared for chromatography with a fine filtration step to remove aggregates formed during TFF. Capture step using CIMmultus QA column is designed in bind-elute mode with salt gradient, to facilitate concentration of virus and removal of impurities. The in-process samples are analyzed using a high-throughput chromatographic approach on PAT fix HPLC system with analytical CIMac Adeno column. The analytical method was developed to track Adenovirus, control Adenovirus harvests, as well as follow the reduction of impurities through the manufacturing processes. Additional feature of such HPLC analytics is multi angle light scattering detector, which serves as orthogonal quantification method. Another important tool is PicoGreen fluorescence detection, very helpful when following the removal of chromatin and DNA residuals, known to cause side effects of vaccines.

During purification method development different materials were tested and compared in terms of purity and virus capacity. Working hand by hand with upstream turned out to be effective way for optimal process development.

List of takeaways: 

• CIMmultus QA is offering effective approach to tackle Adenovirus production bottlenecks, improve purity and quality of Adenovirus vaccines.
• CIMmultus QA columns provide higher yields at lower manufacturing costs and are readily available, overcoming the shortage of raw material supply on the market.
• Accompanying HPLC analytics using CIMac Adeno column is a reliable method offering rapid in-process insight for upstream and downstream applications.
• Additional feature of HPLC analytics is multi angle light scattering detector, which serves as orthogonal quantification method and PicoGreen fluorescence detection, very helpful when following the removal of chromatin and DNA residuals, known to cause side effects of vaccines.
• Robustness of developed process has been confirmed using various USP materials.