Protein A media
for antibody purification


Protein A media is selected as the first step of antibody purification in pharmaceutical company. In this step, it is important to purify high volume of culture supernatant in short time. So it is necessary to select high performance media described in below,

  1. 1. High dynamic binding capacity at high flow rate
  2. 2. Low back pressure at high flow rate
  3. 3. Reusability

Cellufine™ SPA-HC is the resin immobilized alkali resistance protein A ligand. The resin features low back pressure because of its crosslinking technology, and has high porosity in order to achieve high mass transfer of antibody.

The unique beads manufacturing technology obtained through experience of 40th years of business in chromatography makes the resin achieved high dynamic binding capacity at high flow rate.
Cellufine SPA-HC was developed expecting used in manufacturing scale. So current equipment which you have can be availed achieving short of manufacturing time.

Property Characteristics
Ligand Alkali-stable rProtein A affinity ligand produced in E.coli
Matrix Highly cross-linked cellulose beads
Particle size Average 70 µm diameter
Ligand coupling method Coupling via formyl groups on resin
Flow velocity ≧650 cm/h (0.3 MPa) with 10 cmID x 20 cmL Column in water at 24 ℃
Dynamic binding capacity (DBC) ≧ 70 mg /ml (DBC C10 with Polyclonal IgG) 1
Recommend CIP solution 0.1 M NaOH
Temperature stability 4 - 50 ℃ No significant change in performance after 1-week storage.
Storage 2 - 8 ℃ in 20 % (v/v) ethanol
Chemical stability No significant change in performance after 1-week storage in 30 % (v/v) isopropanol, 20 – 70 % (v/v) ethanol, 8 M Urea, 6 M Guanidine-HCl or 0.1 M Acetic acid.
pH working range 3-12


  • Coupled with an alkali resistance rProtein A ligand.
  • Low back pressure due to its crosslinking technology achieving high porosity to facilitate high mass transfer of antibody with the immobilized affinity ligand.
  • A unique smaller bead diameter manufacturing technology producing a resin with high dynamic binding capacity at high flow rates.

Cellufine SPA-HC has been developed for use with current equipment to achieve higher throughput and shorter manufacturing processes to accelerate production of antibody based biologics.

Excellent dynamic binding capacity

Cellufine SPA-HC shows high dynamic binding capacity with a human mAb sub-type 1 antibody (see Figure.1) indicating that this new resin can be used for efficient mAb purification workflows.

Measurement data of dynamic binding capacity of Cellufine SPA-HC, DBC data of polyclonal antibody and monochrome antibody
Figure1Human pAb (IgG for infusion) was purchased commercially. Human mAb IgG1 was purified from CHO cell culture.

Low back pressure at high flow rate

As Cellufine SPA-HC is based on a highly cross-linked cellulose bead structure and is very adaptable for use in conventional manufacturing with large scale columns. Pressure-flow rate studies indicate that Cellufine SPA-HC developed back-pressures of <0.3MPa at 650 cm/h in a 10cmID x 20 cmL bed height (1.57 L) column.

The above data suggests that Cellufine SPA HC resin packed in a pilot process column can be operated at a residence time of 2.4min at < 0.3MPa with a flow rate of 650 cm/h and achieve a mAb capture loading of 50 mg/mL from a clarified CHO cell sample.

Flow property of Cellufine SPA-HC
Figure2Pressure/Flow test was carried out in a 10 cmID x 20 cmL column (Axichrom 10/300, 1.57L CV, GE healthcare) with a water mobile phase at 20-25 oC.

Robust performance not depend the sort of monoclonal antibody

Monoclonal antibodies with a range of isoelectric points (pI) and sub-types were used to investigate whether the molecular charge state had any impact on the dynamic binding capacity of the resin. The results indicate that Cellufine SPA-HC can adsorb antibodies with a wide range of pI’s and show higher DBC capacity compared to an 85 µM high capacity agarose competitive resin. A summary is shown in Figure 3, below.

Adsorption capacity of different monochrome antibodies with Selfine SPA-HC
Robust performance not depend the sort of monoclonal antibodyMonoclonal antibody was used various isoelectric point including humanized antibody divertive.

Excellent reusability

Cleaning in place (CIP) with base is routinely applied during cycles of re-use of rProtein affinity resins. Cellufine SPA-HC has been evaluated for 0.1 M NaOH base CIP for up to 150 cycles of re-use. Results are summarized in Figure 4 below.
The data shows high retention (>95%) of DBC after 150 cycle 0.1M NaOH CIP. The capacity at cycle zero was 75 mg/mL and changed to 72 mg/mL at the end of the study.

Repeated use data of Cellufine SPA-HC, clean-in-place with 0.1M NaOH
Figure4cleaning in place study with 0.1 M sodium hydroxide contact at 15 min.
This figure shows remaining dynamic binding capacity as a ratio. 10 % breakthrough DBC was 76 mg/mL at the first time. After 150 cycle alkaline attachment, DBC changed 72 mg/mL adsorption. It means that 95% activity was remaining after this study finished.

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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