Conjugation and radiolabeling of monoclonal antibodies with zirconium-89 for PET imaging using the bifunctional chelate p-isothiocyanatobenzyl-desferrioxamine

Nat Protoc. 2010 Apr;5(4):739-43. doi: 10.1038/nprot.2010.13. Epub 2010 Mar 25.

Abstract

The positron emitter zirconium-89 ((89)Zr) has very attractive properties for positron emission tomography (PET) imaging of intact monoclonal antibodies (mAbs) using immuno-PET. This protocol describes the step-by-step procedure for the facile radiolabeling of mAbs or other proteins with (89)Zr using p-isothiocyanatobenzyl-desferrioxamine (Df-Bz-NCS). First, Df-Bz-NCS is coupled to the lysine-NH(2) groups of a mAb at pH 9.0 (pre-modification), followed by purification using gel filtration. Next, the pre-modified mAb is labeled at room temperature by the addition of [(89)Zr]Zr-oxalic acid solution followed by purification using gel filtration. The entire process of pre-modification, radiolabeling and purification steps will take about 2.5 h.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal* / chemistry
  • Chelating Agents
  • Deferoxamine / analogs & derivatives
  • Head and Neck Neoplasms / diagnostic imaging
  • Humans
  • Isothiocyanates
  • Mice
  • Mice, Nude
  • Positron-Emission Tomography / methods*
  • Radioisotopes
  • Radiopharmaceuticals / chemical synthesis*
  • Radiopharmaceuticals / chemistry
  • Transplantation, Heterologous
  • Zirconium*

Substances

  • 4-isothiocyanatobenzyl-desferrioxamine
  • Antibodies, Monoclonal
  • Chelating Agents
  • Isothiocyanates
  • Radioisotopes
  • Radiopharmaceuticals
  • Zirconium
  • Deferoxamine