Identification of polyol-responsive monoclonal antibodies for use in immunoaffinity chromatography.
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.
Thompson
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NE
University of Wisconsin, Madison, Wisconsin, USA.
Burgess
R R
RR
D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000937:Antigen-Antibody Reactions / Q000187:drug effects*; D002846:Chromatography, Affinity / Q000379:methods*; D007158:Immunologic Techniques; D011108:Polymers / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D011108:Polymers; C024617:polyol