title: 124I-labeled engineered anti-CEA minibodies and diabodies allow high-contrast, antigen-specific small-animal PET imaging of xenografts in athymic mice.
abstract: Prolonged clearance kinetics have hampered the development of intact antibodies as imaging agents, despite their ability to effectively deliver radionuclides to tumor targets in vivo. Genetically engineered antibody fragments display rapid, high-level tumor uptake coupled with rapid clearance from the circulation in the athymic mouse/LS174T xenograft model. The anticarcinoembryonic antigen (CEA) T84.66 minibody (single-chain Fv fragment [scFv]-C(H)3 dimer, 80 kDa) and T84.66 diabody (noncovalent dimer of scFv, 55 kDa) exhibit pharmacokinetics favorable for radioimmunoimaging. The present work evaluated the minibody or diabody labeled with (124)I, for imaging tumor-bearing mice using a high-resolution small-animal PET system.
  
  
  METHODS
  Labeling was conducted with 0.2-0.3 mg of protein and 65-98 MBq (1.7-2.6 mCi) of (124)I using an iodination reagent. Radiolabeling efficiencies ranged from 33% to 88%, and immunoreactivity was 42% (diabody) or >90% (minibody). In vivo distribution was evaluated in athymic mice bearing paired LS174T human colon carcinoma (CEA-positive) and C6 rat glioma (CEA-negative) xenografts. Mice were injected via the tail vein with 1.9-3.1 MBq (53-85 microCi) of (124)I-minibody or with 3.1 MBq (85 microCi) of (124)I-diabody and imaged at 4 and 18 h by PET. Some mice were also imaged using (18)F-FDG 2 d before imaging with (124)I-minibody.
  
  
  RESULTS
  PET images using (124)I-labeled minibody or diabody showed specific localization to the CEA-positive xenografts and relatively low activity elsewhere in the mice, particularly by 18 h. Target-to-background ratios for the LS174T tumors versus soft tissues using (124)I-minibody were 3.05 at 4 h and 11.03 at 18 h. Similar values were obtained for the (124)I-diabody (3.95 at 4 h and 10.93 at 18 h). These results were confirmed by direct counting of tissues after the final imaging. Marked reduction of normal tissue activity, especially in the abdominal region, resulted in high-contrast images at 18 h for the (124)I-anti-CEA diabody. CEA-positive tumors as small as 11 mg (<3 mm in diameter) could be imaged, and (124)I-anti-CEA minibodies, compared with (18)F-FDG, demonstrated highly specific localization.
  
  
  CONCLUSIONS
  (124)I labeling of engineered antibody fragments provides a promising new class of tumor-specific probes for PET imaging of tumors and metastases.
authors.affiliation: Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at UCLA, Los Angeles, California 90095, USA.
authors.forename: Gobalakrishnan
authors.initials: G
authors.lastname: Sundaresan
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Yazaki
authors.forename: John E
authors.initials: JE
authors.lastname: Shively
authors.forename: Ronald D
authors.initials: RD
authors.lastname: Finn
authors.forename: Steven M
authors.initials: SM
authors.lastname: Larson
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Raubitschek
authors.forename: Lawrence E
authors.initials: LE
authors.lastname: Williams
authors.forename: Arion F
authors.initials: AF
authors.lastname: Chatziioannou
authors.forename: Sanjiv S
authors.initials: SS
authors.lastname: Gambhir
authors.forename: Anna M
authors.initials: AM
authors.lastname: Wu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000493:pharmacokinetics; D000941:Antigens / Q000378:metabolism*; D002272:Carcinoembryonic Antigen / Q000378:metabolism*; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000000981:diagnostic imaging* / Q000378:metabolism; D005260:Female; D005910:Glioma / Q000000981:diagnostic imaging*; D006801:Humans; D018796:Immunoconjugates / Q000493:pharmacokinetics; D007457:Iodine Radioisotopes / Q000493:pharmacokinetics; D007553:Isotope Labeling / Q000379:methods; D008657:Metabolic Clearance Rate; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D015202:Protein Engineering / Q000379:methods; D011786:Quality Control; D011863:Radioimmunoassay / Q000379:methods; D019275:Radiopharmaceuticals / Q000493:pharmacokinetics; D014018:Tissue Distribution; D014055:Tomography, Emission-Computed / Q000379:methods*; D014183:Transplantation, Heterologous
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D002272:Carcinoembryonic Antigen; D018796:Immunoconjugates; D007457:Iodine Radioisotopes; D019275:Radiopharmaceuticals