Antigen binding chromatography

In addition to the covalent binding, some methods derived from bioaffinity chromatography can be used for non covalent attachment of antibodies to a surface by the inactive Fc portion. The advantage is that antigen binding sites stay undamaged and accessible for the analytes due to the orientation of antibody with the active Fab portions towards the tested medium. 

Affinity chromatography is a particularly powerful procedure that can be used to purify IgG, subpopulations of IgG, or the antigen binding fraction of IgG present in serum/ascitic fluid/hybridoma culture supernatant. This technique requires the production of a solid matrix to which a ligand, which either has affinity for the relevant IgG or vice versa, has been bound. Examples of ligands useful in this context are ... 

Clezardin, P., McGregorJ. L., Manach, M.,Boukerche, H., arid Dechavanne, M (1985) One-step procedure for the rapid isolatiort of mouse monoclonal antibodies and their antigen binding fragments by fast protein liquid chromatography on a Mono-Q anion exchange column./ Chromatogmphy S19, b - . 

Fractogel-DEAE chromatography 158 Fragment antigen binding 1088 Fragmin 457 Fraxiparine 457 Fraxodi 457... 

Antigen affinity chromatography Useful for all types of antibody isolation Binds specific antibodies Must make specific antigen-linked column/matrix... 

Riordan, and M. Whitlow (1988). Single-chain antigen-binding proteins. Science 242 423-426. Blank, K., R Lindner, B. Diefenbach, and A. Pluckthun (2002). Self-immobilizing recombinant antibody fragments for immunoaffinity chromatography generic, parallel, and scalable protein purification. Protein Expr. Purif. 24 313-322. 

Immunoaffinity chromatography utilizes the high specificity of antigen—antibody interactions to achieve a separation. The procedure typically involves the binding, to a soHd phase, of a mouse monoclonal antibody which reacts either directly with the protein to be purified or with a closely associated protein which itself binds the product protein. The former approach has been appHed in the preparation of Factor VIII (43) and Factor IX (61) concentrates. The latter method has been used in the preparation of Factor VIII (42) by immobilization of a monoclonal antibody to von WiHebrand factor [109319-16-6] (62), a protein to which Factor VIII binds noncovalenfly. Further purification is necessary downstream of the immunoaffinity step to remove... 

Affinity chromatography, which is the binding of bio-molecules with the matrix bed, often used for antibodies and antigens... 

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