Cellufine Phenyl EX
Flow-through purification of antibody aggregates

Cellufine Phenyl EX

Cellufine Phenyl EX is a chromatography resin that can efficiently remove aggregates of antibody drugs using our unique ligand modification technology. The antibody aggregates are captured to the column, but most of the antibody monomers are recovered as they are in the flow-through fraction. Cellufine Phenyl EX can be used for flow-through purification that selectively adsorbs such as aggregates.

Grade Cellufine Phneyl EX Cellufine MAX Phenyl Cellufine MAX Phenyl LS
Base Cross-linked cellulose Highly cross-linked cellulose Highly cross-linked cellulose
Particle diameter ca. 40~130 μm
Ligand Phenyl group
BSA adsorption capacity [mg/ml] ≥ 13 ≥ 14 ≥ 6
BSA recovery [%] > 30 > 40 > 90
Operating pressure < 0.2 MPa > 0.3 MPa > 0.3 MPa
pH stability pH 2 - 13
Supplied Suspension in 20 % EtOH

Partial structure

  • Cellufine Phenyl EX


    Ligand structure of Cellufine Phenyl EX, phenyl group immobilized on spherical porous cellulose particles

Model Protein Separation

The optimum amount of ligand for HIC resins vary depending on the application. Below figure shows model protein separation with Cellufine Phenyl EX and Cellufine MAX Phenyl. Protein separation studies show that relative binding strength are Phenyl EX > MAX Phenyl. The strength of hydrophobicity is also Cellfine Phenyl EX > Cellfine MAX Phenyl.

Model Proteins Separation
6.6 mm ID x 50 mm L
Buffer A
10 mM Phosphate buffer, pH 7
Buffer B
10 mM Phosphate buffer, pH 7 + 1.5 M Ammonium sulphate
Ribonuclease A, α-chymotrypsinogen A, Lysozyme

Purification of antibody aggregates

Cellufine Phenyl EX is the best chromatography resin for removing aggregates from monoclonal antibodies in flow-through mode. Antibody aggregates were removed using the monoclonal antibody purified with a protein A column. Cellufine Phenyl EX can be worked with low electrical conductivity and has high agglomerate removal properties. In this study, the conductivity of the sample was adjusted to 6 mS / cm. For this reason, it is possible to suppress corrosion of pipes and buffer precipitation due to high salt concentration, which is used with ordinary HIC resins.

Removal of aggregates from monoclonal antibodies
1 mL mini column
Residence time 4 min (75cm/h)
mAb after protein A column 6.6 mg/mL, pH6, 6 mS/cm
Load antibody
93 mg_mab/mL_CV
Equilibration and wash
20 mM AcOH-Tris + NaCl, pH6, 6 mS/cm

As shown in Table, the monoclonal antibody aggregate was present at 3.6% before column loading, but it was reduced to 0.4% by passing Cellufine Phenyl EX.

Resin Aggregate (Before load)[%] Aggregate (After load)[%] Recovery [%]
Cellufine Phenyl EX 3.6 0.4 87
Cellufine MAX Phenyl 3.6 1.3 99

Flow Property

Cellufine Phenyl EX enable high-flow operation, which is essential to efficient purification of biopharmaceuticals. Cellufine Phenyl EX is operable at practical flow velocities and pressures.

Pressure-flow property of Cellufine Phenyl EX
2.6cm I.D. x 19.3 cm L
23 - 25 °C
Mobile phase
Pure water
Resin was packed with a compression factor 1.35. System pressure was excluded from the data.

Repeat use

Cellufine Phenyl EX can be used repeatedly. For cleaning-in-place, use a cleaning solution containing 0.5 M sodium hydroxide and 30% isopropanol. By performing cleaning-in-place with an appropriate cleaning solution, the adsorption performance did not change even after repeated use 60 times.

Performance after repeated use
1 mL Mini-Column
Human gamma globulin 2 mg/mL, in equilibration buffer
20 mM Phosphate buffer, 0.5 M Ammonium Sulphate, pH7.0
20 mM Phosphate buffer, pH7.0
0.5M NaOH+30% IPA(10 CV、Exposure time: 20 min)

Chemical Stability

Usable chemicals

  • Ethanol (70%)
  • Sodium hydroxide (0.5M)
  • Isopropyl alcohol (30%)
  • Detergents
  • Guanidine hydrochloride (6M)
  • Autoclave (121 ºC, 20 min)
  • Urea (6M)

Custom Resin Development Services

JNC offers a custom resin development service for optimization of current products to meet stringent or enhanced performance working directly with large and small customers. In addition, JNC also offers custom resin development services to support new applications of JNC resins.

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