Project Description

Protein A affinity chromatography medium by EPRUI is based on the monodisperse porous poly (methyl methacrylate) (PMMA) microspheres as the matrix, and genetically modified protein A as the ligand, which ensures its high affinity binding of monoclonal antibodies and Fc fragment containing recombinant proteins (macromolecules). Our protein A chromatography media has high mechanical strength and good pH stability. Even at a high flow rate, the beads still maintained a high dynamic binding capacity, meeting the demands from laboratory preparation to industrial production.

1. Highly uniform particle size
EPRUI protein A affinity chromatography medium has highly uniform particle size distribution and precisely controlled pore structure, as shown in Fig. 1.

highly-uniform-particle

 

2. High flow rate
EPRUI protein A affinity chromatography media is made of polymethyl methacrylate (PMMA) microspheres. Our protein A media is able to withstand pressure of 0.5 MPa. Compared with the affinity medium with conventional alagarose matrix, EPRUI protein A will make the separation process much faster, therefore reducing the purification time. It is ideal chromatography media for hundreds of liters of large scale chromatography preparative column.
Experimental:Column:4.6 mm × 50 mm   Mobile phase:50 mM PBS,0.5 M NaCl
flow-rate
Figure 2. Comparison of EPRUI protein A media and a conventional agarose type protein A medium pressure vs. linear flow rate.

3. Excellent loading capacity
At a high flow rate, EPRUI protein A media shows a much better dynamic binding capacity (DBC) than that of the conventional agarose type protein A medium. Even when the residence time was increase to 4-6 minutes.(Experimental:Column:7 mm × 25 mm   Sample:Human IgG,2 mg/mL    Equilibration buffer:20 mM PBS,pH 7.0 ,0.15 M NaCl  Elution:0.1 M Glycine ,pH 3.0)
loading-capacity
Figure3. Comparison of EPRUI protein A media with a leading brand of agarose type Protein A medium for their dynamic binding capacity

4. Outstanding pH stability
EPRUI protein A affinity chromatography medium could be applied in a pH range from 3-12, and it could be cleaned with 0.1-0.5 M NaOH. In a life-cycle test,EPRUI protein A medium remains more than 90% of the original dynamic binding capacity (DBC), after 100 CIP cycles with 0.5 M NaOH as CIP reagent.  (CIP procedure:Medium:Uni®Mab Column:7 mm × 25 mm   Sample:recombinant antibody(chimera)    DBC calculation:10% breakthrough   Equilibration buffer:20 mM PBS,150 mM NaCl, pH 7.2,3 CV(column volume)   Elution:20 mM Citrate,pH 3.0  CIP:0.5 M NaOH,15 CV  Re-equilibration:10 CV)
life-cycle

Figure 4.EPRUI protein A media pH stability in life cycle tests


FeaturesAdvantages
Highly uniform particle sizeGood reproducibility ,easy to pack
Good mechanical strengthHigher pressure resistance, able to pack preparative column in large scale
Higher flow rateHigher productivity
High loading capacityCost effective
Better alkaline stabilityClean with 0.1-0.5M NaOH

EPRUI Protein A media was successfully applied in purification of hIgG from human serum.

Experimental:
Column: 7 mm×25 mm
Sample: human serum, 5 mL
Equilibration Buffer: 20mM PBS, pH 7.0  0.15 M NaCl
Elution: 0.1 M Glycine, pH 3.0
Residence time: 4 min

eprui-protein-a-media-used-in-hIgg-purification-from-human-serum


MatrixMonodisperse PMMA Microspheres
Diameter50um
LigandRecombinant Protein A
hIgG Dynamic Binding Capacity>35mg/ml(4 min residence time)
Linear Flow Rate300-700cm/h
Maximum Pressure72.5 psi(5 bar,0.5Mpa)
PH Stability3-12
Temperature4-40℃
CIP Reagent0.1-0.5M NaOH
Storage20% ethanol, 2-8℃

Prepacked Column Specification:

Dimension7mm*25mm
CartridgePolypropylene(PP)
Column volume1ml
hIgG Dynamic Binding Capacity>35mg/ml(4 min residence time)
Flow Rate0.25-1.0ml/min
Maximum Pressure43.5 psi(3 bar, 0.3Mpa)