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

Claude F. Meares, Michael J. McCall, Dayton T. Reardan, David A. Goodwin, Carol I. Diamanti, Maureen McTigue

Research output: Contribution to journalArticle

348 Scopus citations

Abstract

Preparation of the chelating agent (S)-4-[2,3-bis[bis(carboxymethyl)amino]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.

Original languageEnglish (US)
Pages (from-to)68-78
Number of pages11
JournalAnalytical Biochemistry
Volume142
Issue number1
DOIs
StatePublished - 1984

Keywords

  • gel filtration
  • immunoglobulins
  • luminescence
  • metal ions
  • organic synthesis
  • radioactive
  • radioactivity measurement
  • thin-layer chromatography

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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