General description
Protein A chromatography, the most widely used type of affinity chromatography, relies on the specific and reversible binding of antibodies to an immobilized ligand; in this case protein A. Protein A is a 56 kDa surface protein native to the cell wall of the bacterium Staphylococcus aureus. It is composed of five immunoglobulin-binding domains, each of which are able to bind proteins from many mammalian species, most notably Immunoglobulin G (IgG) through the heavy chain within the Fc region. While the native form of Protein A was used as the ligand for first generation Protein A resins, the recombinant form (rProtein A) produced in E. coli is the most prevalent today. Modifications to the protein structure of the ligand, the advent of ligands composed of single domain multimers, and multipoint attachment have given rise to the caustic stable, high capacity and extremely robust Protein A resins in use today.The protein A ligand can either bind directly to the Fc region of an antibody or to an Fc tag that has been fused to the target of interest. Protein A chromatography is a very robust purification procedure and is used as a capture step due to its specificity and, depending on the intended use for the target molecule (antibodies for diagnostic testing), might be the only chromatographic step required to achieve adequate product purity.Toyopearl AF-rProtein A-650F utilizes a new alkaline resistant recombinant Protein A variant. This new non-animal derived ligand is bound to the base bead, Toyopearl HW-65F, using multipoint attachment. This contributes to both the new resin′s alkaline resistance and its low ligand leakages in the range 5-25ng/mg IgG.Toyopearl AF-rProtein A-650F maintains its capacity well after repeated CIP exposure to 0.1mol/L NaOH and has reasonable tolerance to 0.5mol/L NaOH CIP.The new resin also tolerates exposure to above ambient temperatures and if needed can be autoclaved.Good pressure-flow characteristics and kinetics contribute to maintaining good capacity particularly at low residence times.
Legal Information
Toyopearl is a registered trademark of Tosoh Corporation
Physical form
Shipped in 20% (v/v) ethanol.
72% (v/v) slurry in 20% (v/v) ethanol
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Specifications
Clean in place with 0.1 M NaOH.
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