Pierce™ Protein A/G Agarose

Pierce Protein A/G Agarose consists of purified Protein A/G recombinant fusion protein that has been covalently immobilized onto high-quality crosslinked 6% beaded agarose (CL-6B). It provides the most versatile combination of chromatographic features for high yield and high purity purification of whole IgG from mammalian serum samples. The agarose beads have physical and chemical properties suitable for many affinity purification systems.

Features of Pierce Protein AG Agarose:
• Protein A/G—immobilized recombinant fusion protein of the antibody-binding domains of Protein A and Protein G for polyclonal IgG purification from nearly any mammalian species
• Agarose resin—crosslinked 6% beaded agarose (CL-6B) support, the most popular resin for protein affinity purification methods
• Inert and stable—superior manufacturing method immobilizes Protein A/G by charge-free, leach-resistant covalent bonds, resulting in low nonspecific binding and enabling multiple uses without decline in yield
• Standard capacity—has a normal load of immobilized Protein A/G, providing a binding capacity greater than 7 mg human IgG/mL resin

Protein A/G is a genetically engineered protein that combines the IgG binding domains of both Protein A and Protein G. The fusion protein is expressed in E. coli. Protein A/G contains four Fc binding domains from Protein A and two from Protein G, resulting in a final mass of 50,460 Daltons (40 to 45kDa by SDS-PAGE). Protein A/G is not as pH dependent as Protein A alone but has the additive properties of Protein A and G.

Protein A/G binds to all mammalian IgG subclasses, making it the ideal choice for purifying polyclonal or monoclonal IgG antibodies whose subclass identities have not been determined. The Protein A/G ligand can bind with IgA, IgE, IgM, and to a lesser extent, IgD. Moreover, it has a strong affinity towards all mouse IgG subclasses. However, it cannot bind with mouse IgA, IgM, or serum albumin. This makes Protein A/G an excellent tool for purification and detecting mouse monoclonal antibodies from IgG subclasses without interference from IgA, IgM, and murine serum albumin.

Because the immobilized Protein A/G combines the immunoglobulin-binding domains of both Protein A and Protein G, the individual subclasses of mouse monoclonals are likely to have a stronger affinity to the chimeric Protein AG than to either Protein A or Protein G.

Pierce Protein A/G Agarose is prepared using Thermo Scientific AminoLink coupling chemistry, which provides several advantages compared to traditional methods of ligand immobilization. AminoLink immobilization results in conjugation between sugar monomers of the agarose beads and native lysine residues on the Protein A via simple amide bonds. Unlike typical cyanogen bromide (CNBr) immobilization, the AminoLink method does not introduce any novel chemical groups that could cause undesired nonspecific binding and produces a stable, essentially irreversible bond. The result is a high-binding-capacity resin that retains functional immobilized Protein AG through multiple rounds of antibody purification.

Properties of crosslinked 6% beaded agarose (CL-6B):
• pH Stability: 2 to 14 (short term); 3 to 13 (long term)
• Average particle size: 45 to 165 microns
• Exclusion limit: 10,000 to 4,000,000 Daltons
• Maximum volumetric flow rate: approx. 1 mL/minute (for 1 cm diameter column)
• Maximum linear velocity: 30 cm per hour
• Maximum pressure: less than 25psi (1.5 bar), defined as the maximum pressure drop across a column that the resin can withstand (Note: The indicated gauge pressure of a liquid chromatography apparatus may be measuring the total system pressure rather than the pressure drop across the column.)

More product data
Using antibody-binding proteins for antibody purification

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