The bioconjugation and radiosynthesis of 89Zr-DFO-labeled antibodies

J Vis Exp. 2015 Feb 12:(96):52521. doi: 10.3791/52521.

Abstract

The exceptional affinity, specificity, and selectivity of antibodies make them extraordinarily attractive vectors for tumor-targeted PET radiopharmaceuticals. Due to their multi-day biological half-life, antibodies must be labeled with positron-emitting radionuclides with relatively long physical decay half-lives. Traditionally, the positron-emitting isotopes 124I (t1/2=4.18 d), 86Y (t1/2=14.7 hr), and 64Cu (t1/2=12.7 hr) have been used to label antibodies for PET imaging. More recently, however, the field has witnessed a dramatic increase in the use of the positron-emitting radiometal 89Zr in antibody-based PET imaging agents. 89Zr is a nearly ideal radioisotope for PET imaging with immunoconjugates, as it possesses a physical half-life (t1/2=78.4 hr) that is compatible with the in vivo pharmacokinetics of antibodies and emits a relatively low energy positron that produces high resolution images. Furthermore, antibodies can be straightforwardly labeled with 89Zr using the siderophore-derived chelator desferrioxamine (DFO). In this protocol, the prostate-specific membrane antigen targeting antibody J591 will be used as a model system to illustrate (1) the bioconjugation of the bifunctional chelator DFO-isothiocyanate to an antibody, (2) the radiosynthesis and purification of a 89Zr-DFO-mAb radioimmunoconjugate, and (3) in vivo PET imaging with an 89Zr-DFO-mAb radioimmunoconjugate in a murine model of cancer.

Publication types

  • Video-Audio Media

MeSH terms

  • Animals
  • Antibodies, Monoclonal / chemistry*
  • Cell Line, Tumor
  • Deferoxamine / chemistry
  • Heterografts
  • Humans
  • Immunoconjugates / chemistry
  • Male
  • Mice
  • Mice, Nude
  • Positron-Emission Tomography / methods
  • Prostatic Neoplasms / diagnostic imaging
  • Radioisotopes / chemistry*
  • Radiopharmaceuticals / chemical synthesis
  • Radiopharmaceuticals / chemistry*
  • Zirconium / chemistry*

Substances

  • Antibodies, Monoclonal
  • Immunoconjugates
  • Radioisotopes
  • Radiopharmaceuticals
  • Zirconium
  • Deferoxamine