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  • Name: Protein A Media
  • Id: AE001


Uni®Mab is a high performance Protein A affinity chromatography medium, introduced by Nano-Micro with its patented technology developed in-house. This medium is based on the monodisperse porous poly (methyl methacrylate) (PMMA) microspheres as the matrix, and a genetically modified protein A as the ligand, which ensures its high affinity binding of monoclonal antibodies and Fc fragment containing recombinant proteins (macromolecules). Uni®Mab has come with high mechanical strength and good pH stability. Even at a high flow rate, it still maintained a high dynamic binding capacity, meeting the demands from laboratory preparationto industrial production.

 

Characteristics and Advantages

 

Highly uniform particles
Uni®Mab affinity chromatography medium has highly uniform particle size distribution and precisely controlled porestructure, as shown in Fig. 1.

Figure 1. SEM diagram of Uni®Mab and a leading brand of protein A medium

 

High flow rate
Uni®Mab affinity chromatography medium was made from polymethyl methacrylate (PMMA) microspheres (as matrix); and it would withstand the pressure of 0.5 MPa. When compared to an affinity medium with convention alagarose matrix of the chromatography media, it would make the separation process much faster, thus reducing the time purification, for hundreds of liters of large scale chromatography preparative column.(ExperimentalColumn4.6 mm × 50 mm   Mobile phase50 mM PBS0.5 M NaCl)

Figure 2. Comparison of Uni®Mab and a conventional agarose type protein A medium, their pressure vs. linear flow rate

 

Excellent loading capacity
At a high flow rate, Uni®Mab showed a significantly better dynamic binding capacity (DBC) than that of the conventional agarose type protein A medium.  The same observation was also valid, even when the residence time was increase to 4 - 6 minute. (ExperimentalColumn7 mm × 25 mm  SampleHuman IgG2 mg/mL Equilibration buffer20 mM PBSpH 7.0 0.15 M NaCl   Elution0.1 M Glycine, pH 3.0)

Figure 3. Comparison of Uni®Mab with a leading brand of Protein A medium, for their dynamic binding capacity

 

Outstanding pH stability
Uni®Mab 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, Uni®Mab medium remained greater 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 Column7 mm × 25 mm   Sample:recombinant antibody(chimera) DBC calculation:10% breakthrough Equilibration buffer20 mM PBS150 mM NaCl, pH 7.23 CVcolumn volume)   Elution20 mM CitratepH 3.0  CIP:0.5 M NaOH15 CV  Re-equilibration:10 CV)

Figure 4. Uni®Mab pH stability in life cycle tests

 

Application

Uni®Mab was secessfully applied in purification of hIgG from human serum. (ExperimentalColumn7 mm× 25 mm  Samplehuman serum, 5 mL  Equilibration Buffer20mM PBS, pH 7.0  0.15 M NaCl   Elution0.1 M Glycine, pH 3.0  Residence time4 min)

Figure 5.Uni®Mab used in hIgG purification from human serum

 

Technical information


 

Pre-packed column cartridge

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