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Protein A Agarose for the Elution of Antibodies under Mild Conditions
Protein A is a bacterial cell wall component from Staphylococcus aureus1) that specifically binds to the Fc region of IgG derived from various species including human, rabbit, mouse and cow.2) Protein A agarose in which protein A bound to an agarose resin by a covalent coupling method can be used in antibody purification and immunoprecipitation. Antibody purification using protein A agarose usually requires an acidic buffer solution between pH 2.5 and pH 3.0 during elution steps. However, this frequently causes the antibody to undergo acid denaturation, changing its higher-order structure, resulting in antibody aggregation and inactivation. TCI’s protein A agarose (1) uses a genetically modified protein A mutant which allows for the elution of antibodies under mild conditions (pH 4.0), which most antibodies do not denature, as shown in Figure 1.
Figure 1. Purification of human IgG using 1. Most antibodies are eluted at pH 4.0.
Figure 2. Purification of human IgG using competitor’s product.
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References
- 1) Use of staphylococcal protein A as an immunological reagent
- 2) Staphylococcal protein A consists of five IgG-binding domains
