Cellufine™ MAX 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
- Cellufine™ MAX DexS-HbP, DexS-VirS
