Pierce™ Recombinant Protein A Agarose (Thermo Scientific™)

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.

Features of Recombinant Protein A Agarose:

• 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 17 mg human IgG per milliliter of resin
• Agarose resin – crosslinked 6% beaded agarose for small column or batch affinity purification methods

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.

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. 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.)

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