Identification of Polyol‐Responsive Monoclonal Antibodies for Use in Immunoaffinity Chromatography

Abstract

Abstract One of the limitations of immunoaffinity chromatography as been that high‐affinity antigen‐antibody complexes are difficult to dissociate, often leading to inactivation of the protein product during elution from the immobilized antibody. As described in this unit, some antigen‐antibody complexes can be dissociated in the presence of a combination of a low‐molecular‐weight polyhydroxylated compound (polyol) and a nonchaotropic salt. These conditions seem to be generally nondenaturing and, in some cases, even protein‐stabilizing. This type of antibody is designated “polyol‐responsive.“ These antibodies can be easily identified and isolated as monoclonal antibodies (MAbs) from a typical fusion, using standard hybridoma procedures. They have proven to be very valuable reagents for the immunoaffinity purification of active, labile, multi‐subunit protein complexes.