Thermo Scientific Pierce Recombinant Protein A Agarose is a beaded agarose resin to which a genetically engineered form of Protein A has been immobilized for use in IgG purification applications where the native ligand has undesirable effects.
The Pierce Recombinant Protein A Agarose uses a genetically engineered form of Protein A that is produced in E. coli and then immobilized on crosslinked 6% beaded agarose (CL-6B). Although some nonessential regions have been removed, recombinant is similar to native Protein A in overall size and IgG-binding domains. The protein has four high affinity binding sites capable of specifically binding to the Fc region of immunoglobulin molecules from several species.
- Immobilized Protein A – specifically binds IgG from several mammalian species, especially most subtypes of human and rabbit IgG
- Standard capacity – loading of immobilized Protein A provides a binding capacity of 15 to 17mg human IgG per milliliter of resin
- Agarose resin – crosslinked 6% beaded agarose for small column or batch affinity purification methods
Recombinant Protein A is produced in E. coli and functions essentially the same as native Protein A from Staphylococcus aureus. The Protein A molecule contains four high-affinity (Ka= 10^8/mol) binding sites capable of interacting with the Fc region from IgG of several species including human and rabbit.
Some researchers prefer to use an affinity resin based on the recombinant form of Protein A in particular applications where practical experience indicates that the different production source (E. coli vs. S. aureus) provides less nonspecific binding or lower levels of endotoxin contamination. Otherwise, native Protein A resins generally provide higher capacity antibody purification.
Properties of crosslinked 6% beaded agarose (CL-6B):
- Support 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. 1mL/minute (for 1cm diameter column)
- Maximum Linear Velocity: 30cm 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.)
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Review of antibody purification methods
All Protein A Resins (including Protein A Agarose and IgG Purification Kits)
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