Selection of monoclonal antibodies for immunoaffinity chromatography: model studies with antibodies against soy bean trypsin inhibitor

J Biochem Biophys Methods. 1991 Jul-Aug;23(1):73-82. doi: 10.1016/0165-022x(91)90052-x.

Abstract

To facilitate selection of monoclonal antibodies for immunoaffinity chromatography, an ELISA screening procedure was developed. The assay is based on the avidin-biotin system and provides a profile of the monoclonal antibody which is based on the binding characteristics of the antigen binding site when exposed to different elution reagents. The elution profiles of 5 monoclonal antibodies to soy bean trypsin inhibitor (SBTI) were determined and for 2 of the antibodies the results obtained in the ELISA were verified using column experiments. The affinity constants were determined for the same 5 monoclonal antibodies and no correlation was seen with the ease of elution. The elution profiles presented here are easily obtained and the results indicate that a general screening procedure for suitable combinations of antibodies and elution conditions can be carried out using an elution ELISA assay when modified as described herein.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / chemistry*
  • Antigen-Antibody Reactions*
  • Binding Sites, Antibody
  • Chromatography, Affinity
  • Enzyme-Linked Immunosorbent Assay
  • Mice
  • Mice, Inbred BALB C
  • Trypsin Inhibitor, Kunitz Soybean / chemistry
  • Trypsin Inhibitor, Kunitz Soybean / immunology*

Substances

  • Antibodies, Monoclonal
  • Trypsin Inhibitor, Kunitz Soybean