Performing a Purification of IgG Antibodies with HiTrap® Protein A HP columns

Extracted from Affinity Chromatography Vol. 1: Antibodies, GE Healthcare, 2016
 

HiTrap® Protein A HP are 1 ml and 5 ml ready-to-use columns prepacked with Protein A Sepharose® High Performance (Figure 3.17). The columns are used for convenient purification of antibodies from cell culture supernatants, serum, and ascites. Purification can be performed using a syringe, pump, or chromatography system. Furthermore, purification capacity can be greatly increased by connecting columns in series.

HiTrap® Protein A HP columns

Fig 3.17. HiTrap® Protein A HP columns are designed for rapid purification of antibodies using a syringe, pump, or chromatography system.

 

Figure 3.18 shows the purification of mouse monoclonal IgG2b from a hybridoma cell culture supernatant using a 1 ml HiTrap® Protein A HP column.

Purification of mouse monoclonal IgG2b from a hybridoma cell culture supernatant

Fig 3.18. (A) Purification of mouse monoclonal IgG2b from cell culture supernatant on HiTrap® Protein A HP, 1 ml column. Purity of mouse IgG2b was confirmed by (B) nonreducing SDS-PAGE on PhastSystem using PhastGel 10-15 (silver stained), and (C) agarose-gel immunodiffusion.

 

 

Sample preparation/Buffer preparation/Purification

Use the same protoocol as for HiTrap® Protein G HP.

Materials

     
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