Unit

UNIT 11.18 Identification of Polyol-Responsive Monoclonal Antibodies for Use in Immunoaffinity Chromatography

  1. Nancy E. Thompson,
  2. Richard R. Burgess

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1118s54

Current Protocols in Molecular Biology

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.

Author Information

  1. University of Wisconsin, Madison, Wisconsin

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: APR 2001

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.