title: Monoclonal antibodies labeled with rhenium-186 using the MAG3 chelate: relationship between the number of chelated groups and biodistribution characteristics.
abstract: Our previous studies on the preparation of 186Re-MAb conjugates for clinical radioimmunotherapy (RIT) were extended with the aim to derive conjugates which have a high Re:MAb molar ratio, are stable in vitro and in vivo, have favorable biodistribution characteristics and can be used together with 99mTc-MAb conjugates as a matched pair in combined radioimmunoscintigraphy/RIT studies.
  
  
  METHODS
  Rhenium and 99mTc-conjugates of intact MAb E48 were prepared according to our previously described multistep procedure using the MAG3 chelate and analyzed by protein mass spectrometry for the number of chelate molecules coupled to the MAb. For biodistribution analysis, tumor-free nude mice were simultaneously injected with 186Re-, 99mTc/99Tc- and/or 125I-labeled E48 IgG and dissected 1-48 hr postinjection.
  
  
  RESULTS
  Rhenium-186-MAb conjugates with up to 20 Re-MAG3 groups per MAb molecule were prepared with an overall radiochemical yield of 40%-60%. The conjugates did not contain empty MAG3 groups and no aggregates were formed. Only conjugates with a 186Re-MAG3:MAb molar ratio higher than 12 demonstrated slightly impaired immunoreactivity to a maximum of 15% decrease at the 20:1 molar ratio. Biodistribution experiments revealed that a proportion of the conjugate became rapidly eliminated from the blood for conjugates with a Re-MAG3:MAb molar ratio higher than 8. In this case, an increased uptake of activity was observed in the liver and intestines. The 99mTc/99Tc-MAb conjugates showed a similar enhanced blood clearance when containing more than eight Tc-MAG3 groups, while dual labeling of MAbs revealed that the in vivo stability of the conjugated Re-MAG3 complex itself does not differ from the corresponding Tc-MAG3 complex.
  
  
  CONCLUSIONS
  With the method described in this study, it is possible to prepare 186Re-MAG3-MAb conjugates that fulfil all the aforementioned criteria for use in clinical RIT. Coupling of too many metal-MAG3 groups to MAbs results in rapid blood clearance. At the same metal-MAG3:MAb molar ratio, 99mTc/99Tc-MAb conjugates show a similar pharmacokinetic behavior as 186Re-MAb conjugates and can thus be used to predict the localization of 186Re-labeled MAbs and make dosimetric predictions in individual patients.
authors.affiliation: Department of Otolaryngology/Head and Neck Surgery, Free University Hospital, Amsterdam, The Netherlands.
authors.forename: F B
authors.initials: FB
authors.lastname: van Gog
authors.forename: G W
authors.initials: GW
authors.lastname: Visser
authors.forename: R
authors.initials: R
authors.lastname: Klok
authors.forename: R
authors.initials: R
authors.lastname: van der Schors
authors.forename: G B
authors.initials: GB
authors.lastname: Snow
authors.forename: G A
authors.initials: GA
authors.lastname: van Dongen
mesh_terms: D000818:Animals; D002294:Carcinoma, Squamous Cell / Q000532:radiotherapy; D006258:Head and Neck Neoplasms / Q000532:radiotherapy; D051379:Mice; D008819:Mice, Nude; D016719:Radioimmunodetection; D016499:Radioimmunotherapy; D011868:Radioisotopes / Q000493:pharmacokinetics / Q000627:therapeutic use; D012211:Rhenium / Q000493:pharmacokinetics / Q000627:therapeutic use; D017263:Technetium Tc 99m Mertiatide / Q000493:pharmacokinetics / Q000627:therapeutic use; D014018:Tissue Distribution
chemical_list: D011868:Radioisotopes; D017263:Technetium Tc 99m Mertiatide; D012211:Rhenium