UNIT 11.18 Identification of Polyol-Responsive Monoclonal Antibodies for Use in Immunoaffinity Chromatography
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Thompson, N. E. and Burgess, R. R. 2001. Identification of Polyol-Responsive Monoclonal Antibodies for Use in Immunoaffinity Chromatography. Current Protocols in Molecular Biology. 54:VI:11.18:11.18.1–11.18.9.
- Published Online: 1 MAY 2001
- Published Print: APR 2001
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.