Affinity Chromatography Media

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.

Cellufine MAX DexS
Figure1Lactoferrin bingeing to MAX DexS-HbP and Heparin immobilized agarose

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.

Cellufine MAX DexS
Figure2
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  

Development of the grade

Development of the grade according to demand of a customer (inquiry)

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