Pierce™ Protein A Chromatography Cartridges, 5 mL (Thermo Scientific™)

The Thermo Scientific™ Pierce™ Chromatography Cartridges Protein A are convenient, ready-to-use pre-packed devices for the isolation and purification of polyclonal and monoclonal antibodies. These cartridges are effective for routine affinity purification of polyclonal and monoclonal IgG from serum, ascites and cell culture supernatant.

Pierce Protein A Plus Agarose consists of purified native Protein A that has been covalently immobilized at high density onto high-quality crosslinked 6% beaded agarose (CL-6B). Among the many available varieties of immobilized Protein A affinity resins, this one 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 A Plus Agarose:

• Native Protein A – immobilized Protein A is ideal for polyclonal IgG purification from human, rabbit, pig, dog or cat serum
• Agarose resin – support is crosslinked 6% beaded agarose (CL-6B), the most popular resin for protein affinity purification methods
• Inert and stable – superior manufacturing method immobilizes Protein A by charge-free, leach-resistant covalent bonds, resulting in low nonspecific binding and enabling multiple uses without decline in yield
• High capacity – this “Plus” variety of Pierce Protein A Agarose has a dense load of immobilized Protein A, providing a binding capacity greater than 34 mg human IgG/mL resin (approx. 16 to 17 mg mouse IgG/mL resin)

Pierce Cartridges are compatible with the major automated liquid-chromatography systems or for manual syringe processing, and attach directly to ÄKTA™ or FPLC Systems without additional connectors. An accessory pack, included with each product, readily adapts cartridges for use with Luer-Lok Syringe Fittings or 1/16” tubing. The Protein A cartridges provide fast, easy and reproducible chromatographic separations, and can be regenerated for multiple uses.

Protein A is a cell wall component produced by several strains of Staphylococcus aureus. The 46.7kDa-protein consists of a single polypeptide chain that is essentially devoid of carbohydrate. Native Protein A has contains four high-affinity (Ka = 10^8/mol) binding sites that are capable of interacting with the Fc region of IgG-class antibodies from selected mammalian species. IgG-binding function is optimal at pH 8.2, but efficient binding also occurs in neutral and physiological buffers (pH 7.0 to 7.6).

Protein A is a bacteria-derived protein that binds with high affinity and specificity to the Fc portion of antibodies, especially those of the IgG class. Compared to its alternative (Protein G), Protein A provides the highest binding capacity for subclasses of IgG from rabbits, pigs, dogs and cats. Protein A also has higher binding capacity for human IgG, except for IgG3, which it binds weakly. Protein A binds weakly to mouse IgG1 and is not recommended for most applications with that antibody isotype.

Pierce Protein A Plus 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 A through multiple rounds of antibody purification.

Properties of crosslinked 6% bead