Pierce™ Protein L Plus Agarose (Thermo Scientific™)

Thermo Scientific Pierce Protein L Plus Agarose is a high-capacity Protein L beaded agarose resin for use in a variety of antibody affinity purification methods.

Features of Protein L Plus Agarose:

• Protein L – immobilized Protein L is ideal for selective purification of human and mouse antibodies that have kappa light chains
• 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 L 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 L Agarose has a dense load of immobilized Protein L, providing a binding capacity of 10 to 20 mg human IgG/mL resin

Pierce Protein L Plus Agarose is a high-capacity affinity resin for purification of mammalian IgG isotypes that contain specific kappa light chains. Protein L is especially suited for isolating mouse IgG VkI and human IgG VkI, VkIII and VkIV monoclonal antibodies. The resin consists of recombinant Protein L immobilized on crosslinked 6% beaded agarose (CL-6B) using a stable, leach-resistant chemistry. The agarose beads can be regenerated and reused multiple times when stored properly.

Protein L is an immunoglobulin-binding (36kDa) protein that was originally derived from the bacteria Peptostreptococcus magnus but is now produced recombinantly in E. coli. Protein L has the unique ability to bind through kappa light chain interactions without interfering with an antibody's antigen-binding site. Thus, Protein L will bind members of all different classes and subclasses of immunoglobulin (IgG, IgM, IgA, IgE and IgD) if they have kappa light chains. Protein L will also bind single-chain variable fragments (scFv) and Fab fragments. Protein L binds kappa types I, III, and IV in human and kappa type I in mouse. Furthermore, Protein L has an isoelectric point (pI) of approx. 4.5 to 4.8.

Pierce Protein L 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 L through multiple rounds of 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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Pierce™ Protein L Agarose