Conjugation of antibodies with bifunctional chelating agents: isothiocyanate and bromoacetamide reagents, methods of analysis, and subsequent addition of metal ions

Anal Biochem. 1984 Oct;142(1):68-78. doi: 10.1016/0003-2697(84)90517-7.

Abstract

Preparation of the chelating agent (S)-4-[2,3-bis[bis(carboxymethyl)am ino]propyl]phenyl isothiocyanate is reported. Procedures for conjugation of this and (S)-N-4-[2,3-bis[bis-(carboxymethyl)amino] propyl]phenyl bromoacetamide to monoclonal antibodies and other proteins are described. The conjugates may be purified quickly by centrifugation through Sephadex G-50. The number of protein-bound chelating groups may be measured by titration with standard 57Co2+, using thin-layer chromatography to monitor binding. The labeled products retain their immunoreactivity, as illustrated by experiments in vivo with chelate-conjugated antibody to mouse I-AK antigen.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Monoclonal*
  • Antibody Affinity
  • B-Lymphocytes / immunology
  • Chelating Agents* / chemical synthesis
  • Chromatography, High Pressure Liquid
  • Chromatography, Thin Layer
  • Cobalt Radioisotopes
  • Cross-Linking Reagents* / chemical synthesis
  • Edetic Acid / analogs & derivatives
  • Edetic Acid / chemical synthesis
  • Indium
  • Lymph Nodes / metabolism
  • Magnetic Resonance Spectroscopy
  • Metals*
  • Mice
  • Mice, Inbred BALB C
  • Protein Binding
  • Radioisotopes
  • Spectrophotometry, Infrared
  • Spectrophotometry, Ultraviolet

Substances

  • Antibodies, Monoclonal
  • Chelating Agents
  • Cobalt Radioisotopes
  • Cross-Linking Reagents
  • Metals
  • Radioisotopes
  • Indium
  • 1-(4-bromoacetamidobenzyl)EDTA
  • 4-(2,3-bis(bis(carboxymethylamino))propyl)phenyl isothiocyanate
  • Edetic Acid