CellufineTMMAX DexS-HbP, DexS-VirS
Cellufine MAX DexS is a new chromatography resin incorporating Dextran sulfate polymer surface modification. This non-animal derived pseudo affinity ligand can be used instead of immobilized Heparin for blood protein fractionation and viral capture.
JNC offers two different Cellufine MAX DexS resins, surface modified with different lengths of Dextran Sulfate polymer; a) DexS-HbP developed for purification of heparin binding proteins. and b) DexS-VirS, for purifying virus and virus like particles especially having affinity for heparin. Both resins have been developed to improve the protein or virus capture efficiency of existing resins by employing a pseudo affinity mimetic polymer surface based on Dextran Sulfate. The cross-linked cellulose base bead has been optimized for high flow applications and is fully compatible up to 0.5 M NaOH for CIP. Resin properties are summarized in Table 1
| Performance Characteristics | ||
|---|---|---|
| Product name | Cellufine MAX DexS-HbP | Cellufine MAX DexS-VirS |
| Ligand | Dextran sulfate | |
| Matrix | Cross-linked cellulose beads | |
| Particle size | 40 -130 μm (ca 90μm） | |
| Sulfur contents | ≥ 36 μmol/mL | ≥ 74 μmol/mL |
| Lysozyme adsorption capacity | ≥ 50 mg/mL | ≥ 56 mg/mL |
| pH stability | 3 - 12 | |
| Operating pressure | < 0.3 MPa | |
Table 1Performance Characteristics of Cellufine MAX DexS resins
Features
Cellufine MAX DexS-HbP is a pseudo affinity ligand based on low molecular weight Dextran sulfate modification. This non-animal derived affinity ligand can be used instead of immobilized Heparin for blood protein fractionation. The cross-linked cellulose base bead has been optimized for high flow applications and is fully compatible up to 0.5 M NaOH for CIP. This new resin has been developed to improve the protein capture efficiency of heparin binding proteins purified from plasma, such as Lactoferrin (Figure 1 below) and Antithrombin III.
Capture of Inactivated Influenza Virus with Cellufine MAX DexS-VirS
Dextran sulfate affinity on cross-linked agarose has been used to capture and purify inactivated influenza virus from allantoic and cell culture samples with a high degree of purity.
Inactivated influenza virus (A/duck/Hokkaido/Vac-2/2007 H7N7) was captured and eluted in a purification workflow to measure 10% DBC. below. The study was carried out with a 5 mmID x 2.5cmL (0.49 mL volume) column in 10 mM Na Phosphate, 120 mM NaCl buffer pH 7.4 at a flow rate of 0.2 mL/min (60 cm/h for a residence time of 2.5 min). Viral adsorption breakthrough was estimated by measuring HA viral activity in collected fractions. The results are summarized in Figure 2, 3 and Table 2. Cellufine MAX DexS-VirS showed superior virus adsorption capacity in comparison with other dextran sulfate coated resin and Cellufine Sulfate.
- Resin
- Cellufine MAX DexS-VirS
- Column
- I.D. 5.0 x 25 mmH (0.5 ml)
- Sample
- Allantoic fluid including inactivated flu virus
- Virus type
- A/duck/Hokkaido/Vac-2/2007(H7N7)
- Flow Rate
- 0.2 ml/min(60 cm/hr, R.T. 2.5 min)
- Equilibration
- 10 mM sodium phosphate, 120 mM sodium chloride, pH 7.4
- Wash
- 10 mM sodium phosphate, 150 mM sodium chloride, pH 7.4
- Elute 1
- 10 mM sodium phosphate, 500 mM sodium chloride, pH 7.4
- Elute 2
- 10 mM sodium phosphate, 2 M sodium chloride, pH 7.4
| Step | HA | Total protein | DNA | |||||
|---|---|---|---|---|---|---|---|---|
| HAU | % | mg | % | ng/HAU | mg | % | pg/HAU | |
| Load | 204,800 | 100 | 6.4 | 100 | 31.3 | 24.7 | 100 | 120.6 |
| Flow-through | 7,760 | 3.8 | 4.1 | 64.2 | 530.7 | － | － | － |
| Elute 1 | 139,520 | 68.1 | 0.4 | 5.9 | 2.7 | 3.0 | 12.0 | 21.3 |
| Elute 2 | 64,160 | 31.3 | 0.1 | 1.5 | 1.5 | 3.4 | 13.9 | 53 |
| Total recovery | 211,440 | 103.2 | 4.6 | 71.6 | 21.7 | － | － | － |
Table 2Recovery of Inactivated Influenza Flu Viral particles with Cellufine MAX DexS-VirS
- CellufineTMMAX DexS-HbP, DexS-VirS
