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Purification method for vaccine virus using affinity chromatography
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A purification method, virus technology, applied in the field of separation and purification of vaccine virus
Pending Publication Date: 2021-07-23
HK INNO N CORP
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In addition, there is a disadvantage that impurities with a charge similar to the virus may be eluted together
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Embodiment 1
[0048] Example 1. Use of Capto with dextran sulfate TM Purification of DeVirS resin
[0049] In Example 1, Capto with dextran sulfate ligand was used TM The DeVirS resin demonstrated the purification yield and impurity removal of the vaccine virus.
[0050] 20 mM sodium phosphate pH 7.5 buffer was used as equilibration solution and wash solution (0M sodium chloride) and an eluting solution was prepared and used with pH 7.5 buffer where sodium chloride would reach 2M in the equilibration solution.
[0051] First, a vaccinia virus sample containing enteroviruses is loaded onto the column, which is then washed by flowing with a wash solution. Next, flow the eluent from 0M to 2M NaCl with a linear concentration gradient, collect the eluate, and then 50 The vaccine virus content is measured, and the impurity content is measured.
[0052] figure 2 with image 3 shows the use of Capto with dextran sulfate TM Results of vaccine virus purification on DeVirS resin.
[0053] ...
Embodiment 2
[0056] Example 2. Purification Using HiTrap Heparin Resin Containing Heparin
[0057] In Example 2, the purification yield and impurity removal rate of vaccine virus were confirmed using HiTrap Heparin resin containing heparin ligand.
[0058] 50 mM Tris-HCl pH 8.0 buffer was used as the equilibration solution and washing solution, and an eluting solution was prepared and used so that sodium chloride would reach 2M in the equilibration solution.
[0059] First, a vaccinia virus sample containing enteroviruses is loaded onto the column, which is then washed by flowing with a wash solution. Next, flow the eluent from 0M to 2M NaCl with a linear concentration gradient, collect the eluate, and then 50 The vaccine virus content is measured, and the impurity content is measured.
[0060] Figure 4 with Figure 5 Results of purification of vaccinia virus using HiTrap Heparin resin containing heparin are shown.
[0061] refer to Figure 4 with Figure 5 , when comparing the a...
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Abstract
The present disclosure relates to separation and purification methods for a vaccine virus using affinity chromatography, and more particularly, to a purification method for a virus capable of obtaining a vaccine virus with a high purity and a high yield using affinity chromatography containing a vaccine virus-affinity resin.
Description
technical field [0001] The present disclosure relates to methods for isolating and purifying vaccine viruses using affinity chromatography, and more particularly, to methods for obtaining vaccine viruses of high purity and high yield using affinity chromatography containing virus affinity resins. Virus isolation and purification methods. Background technique [0002] In vaccine viruses cultured using cells derived from species other than humans as host cells, it is necessary to remove host-derived materials. To remove host-derived material, in the prior art, sugar density gradient centrifugation, size exclusion chromatography or ion exchange chromatography have been used. As commonly used methods in virus purification, these methods are used more than affinity chromatography because these methods can be easily applied regardless of the type of virus. [0003] Sugar density gradient centrifugation is a method of purifying viruses using density differences produced by sugars...
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