Chromatography Purification of Inactive Rabies Virus
Rabies is a preventable viral disease most often transmitted through the bite of a rabid animal. The rabies virus infects the central nervous system of mammals, ultimately causing disease in the brain and death. Rabies virus vaccine is still the most effective way to protect humans and animals from rabies virus infection.
There are plenty of methods for the purification of rabies virus vaccine. For vaccines, there are active virus vaccine, inactive virus vaccine, and live attenuated virus vaccine. Each type of vaccine has its own advantages. The final purification methods are based on the vaccine suppliers. We have tested the purification method with VirCap® AF resin for live attenuated viruses. In this test, we studied the purification method of the inactive virus with VirCap® InertShell.
Test Method
- Column: ID 7.3 mm×100 mm CV: .3 mL
- Flow rate: 0.8 mL/min
- Balanced buffer: 20 mM PB pH6.5-8.0 (include NaCl 0-500 mM) Elution/CIP buffer: 1M NaOH + 30% isopropanol
- Sample: Treat the virus fermentation fluid in 8000 rpm centrifugation. Then dilute it by 1 time with a balance buffer
* Test results will change according to different instrument and operators.
Using SEC-HPLC analysis (TSKgel G3000SWXL) to test the breakthrough solution, eluent, and sampling solution, the chromatogram is shown as Pic 1, Pic 2, Pic 3. In Pic 1, the retention time of the target virus molecule is 10.047 min which isn't found in Pic 3. The peak of the target virus molecule is almost all collected in breakthrough compounds in Pic 3-2. The purification effect of VirCap® InerShell in rabies virus sample test is achieved expected target.
When the pH of the equilibrium buffer increased from 6.5 to 8.0, the peak shape of the flow-through component had no significant difference. The concentration of NaCl in the balanced buffer was adjusted from 0 to 500mM, and there was no significant difference in the peak shape of the flow-through components. These tests gave VirCap® Inert Shell a wide operating condition.
* Test results will change according to different instrument and operators.
