Immunoaffinity chromatography.
The basic procedure of immunoaffinity chromatography (IAC) is described. The insoluble support matrices available for IAC and their activation chemistries, including some of the most recently introduced, are reviewed. Means of selecting the most appropriate monoclonal antibody (MAb) are described, although an empirical approach is still required for the final choice of antibody. Precise methods of running IAC columns are surveyed including the binding, washing, and elution stages, although no precise recommendations can be made particularly for the elution step since this is unique to a particular MAb and antigen. All IAC sorbents lose activity with time through a combination of MAb inactivation and ligand leakage. The relative importance of the two phenomena is discussed, and suggestions are made to minimize the problem along with an indication of the relative stabilities of a range of coupling chemistries. A sample of the proteins purified by IAC is given together with pointers to the future of the technique.
Jack
G W
GW
Division of Biotechnology, Centre for Applied Microbiology and Research, Salisbury, UK.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000941:Antigens / Q000302:isolation & purification; D001709:Biotechnology; D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity / Q000379:methods*; D004355:Drug Stability; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology; D006801:Humans; D008024:Ligands; D008969:Molecular Sequence Data; D011506:Proteins / Q000276:immunology / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D000941:Antigens; D006023:Glycoproteins; D008024:Ligands; D011506:Proteins