A new method for site-specific labelling of the oligosaccharide chain of antibodies: preliminary results

J Nucl Biol Med (1991). 1994 Dec;38(4 Suppl 1):15-7.

Abstract

There are various approaches for improving endoradiotherapy and diagnosis with monoclonal antibodies in nuclear medicine. The known methods of site-specific labelling of biomolecules based either on reactions with sulfhydryl groups or on reactions with aldehyde groups of the oligosaccharide chains effect unwanted alterations of the biomolecules. We present a new method to introduce radioactive halogens into the oligosaccharide chains of an antibody, based on the enzymatic transfer of the labelled synthetic sialic acid derivative CMP-9-deoxy-9-salizoyl-NeuAc. It was first labelled by the iodogen-method (iodine) in yields of more than 90%. Under selected conditions it was possible to obtain di- and trihalogenated products. Then the radioactive sialic acid derivatives were transferred during 90 minutes at room temperature with 2.6-sialyltransferase from rat liver into the oligosaccharide chains of antibodies. It is necessary to use neuraminidase pretreated antibodies with an increased number of binding sites for sialic acid derivatives. Yields of about 55% were obtained for the monoiodinated sialic acid derivative. With this method we present a reasonable alternative reaction of labelled compounds with biomolecules. Studies of the immunoreactivity are now in progress.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / chemistry
  • Cytidine Monophosphate / analogs & derivatives
  • Cytidine Monophosphate / chemistry
  • Humans
  • Indicators and Reagents
  • Iodine Radioisotopes
  • Isotope Labeling / methods*
  • Radioimmunodetection*
  • Rats
  • Sialic Acids / chemistry
  • Urea / analogs & derivatives

Substances

  • Antibodies, Monoclonal
  • CMP-9-deoxy-9-salicyl-N-acetylneuraminic acid
  • Indicators and Reagents
  • Iodine Radioisotopes
  • Sialic Acids
  • 1,3,4,6-tetrachloro-3 alpha,6 alpha-diphenylglycoluril
  • Urea
  • Cytidine Monophosphate