Protein L

Protein L is a 36,000 dalton immunoglobulin-binding protein isolated from the bacteria Peptostreptococcus magnus. Unlike Protein A and Protein G, which bind to the Fc region of immunoglobilins (antibodies), Protein L binds antibodies through light chain interactions. Since no part of the heavy chain is involved in the binding interaction, Protein L binds a wider range of antibody classes than Protein A or G. Protein L binds to representatives of all antibody classes, including IgG, IgM, IgA, IgE and IgD. Single chain variable fragments (ScFv) and Fab fragments also bind to Protein L.

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Despite this wide binding range, Protein L is not a universal antibody-binding protein. Protein L binding is restricted to those antibodies that contain kappa light chains.¹ In humans and mice, most antibody molecules contain kappa (κ) light chains and the remainder have lambda (ι) light chains. Protein L is only effective in binding certain subtypes of kappa light chains. For example, it binds human VκI, VκIII and VκIV subtypes but does not bind the VκII subtype. Binding of mouse immunoglobulins is restricted to those having VκI light chains.¹

Given these specific requirements for effective binding, the main application for immobilized Protein L is purification of monoclonal antibodies from ascites or cell culture supernatant that are known to have the kappa light chain. Protein L is extremely useful for purification of VLκ-containing monoclonal antibodies from culture supernatant because it does not bind bovine immunoglobilins, which are often present in the media as a 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.

Other antibody binding proteins

In addition to Protein L, other immunoglobulin-binding bacterial proteins such as Protein A, Protein G and Protein A/G are all commonly used to purify, immobilize or detect immunoglobulins. Each of these immunoglobulin-binding proteins has a different antibody binding profile in terms of the portion of the antibody that is recognized and the species and type of antibodies it will bind.

References

1. Nilson, B.H., et al.(1993). J. Immunol. Methods164, 33-40.
2. Bjorck, L., et al.(1998). J. Immunol140, 1194-1197.
3. Kastern, W., et al.(1992). J. Biol. Chem.267, 12820-12825.
4. Akerstrom, B., and Bjorck, L. (1989). J. Biol. Chem.264, 19740-19746.