| The Thermo Scientific Pierce Chromatography Cartridges Protein L are convenient, ready-to-use prepacked devices for isolation and purification of immunoglobulin classes IgG, IgM, IgA, IgE and IgD via their kappa light chains.Pierce Protein L Agarose consists of recombinant Protein L that has been covalently immobilized onto high-quality crosslinked 6% beaded agarose (CL-6B). This particular variety of the resin provides the most versatile combination of chromatographic features for high yield and high purity purification of target immunoglobulins. The agarose beads have physical and chemical properties suitable for many affinity purification systems.Pierce Chromatography Cartridges are compatible with the major automated liquid-chromatography systems or for manual syringe processing, and attach directly to ÄKTA or FPLC Systems without additional connectors. An accessory pack, included with each product, readily adapts columns for use with Luer-Lok Syringe Fittings or 1/16” tubing. Protein L cartridges provide fast, easy and reproducible chromatographic separations and can be regenerated for multiple uses.Protein L is an immunoglobulin-binding protein (36kDa) that originates from the bacteria Peptostreptococcus magnus, but is now produced recombinantly in E. coli. Unlike Protein A and Protein G, which bind primarily through Fc regions (i.e., heavy chain) of immunoglobilins, Protein L binds Igs through interactions with the light chains. Because no part of the heavy chain is involved in the binding interaction, Protein L binds a wider range of Ig classes than Protein A or G. Protein L binds to representatives of all classes of Ig, including IgG, IgM, IgA, IgE and IgD. Single chain variable fragments (ScFv) and Fab fragments also bind to Protein L.Despite this wide-ranging binding capability with respect to Ig classes, Protein L is not a universal immunoglobilin-binding protein. Binding of Protein L to immunoglobulins is restricted to those containing kappa light chains (i.e., kappa chain of the VL domain). In humans and mice, kappa light chains predominate. The remaining immunoglobulins have lambda light chains. Furthermore, Protein L is effective in binding only certain subtypes of kappa light chains. For example, it binds human VkI, VkIII and VkIV subtypes but does not bind the VkII subtype. Binding of mouse immunoglobulins is restricted to those having VkI light chains.Given these specific requirements for effective binding, the main application for immobilized Protein L is purification of monoclonal antibodies from ascites or culture supernatant that are known to have the kappa light chain. Protein L is extremely useful for purification of VLk-containing monoclonal antibodies from culture supernatant because it does not bind bovine immunoglobilins, which are present in the media serum supplement. Also, Protein L does not interfere with the antigen-binding site of the antibody, making it useful for immunoprecipitation assays, even using IgM.Pierce Protein L 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.Pierce Protein L Agarose is a versatile 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. |
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