title: In vivo biodistribution, PET imaging, and tumor accumulation of 86Y- and 111In-antimindin/RG-1, engineered antibody fragments in LNCaP tumor-bearing nude mice.
abstract: To optimize in vivo tissue uptake kinetics and clearance of engineered monoclonal antibody (mAb) fragments for radiotherapeutic and radiodiagnostic applications, we compared the biodistribution and tumor localization of four (111)In- and (86)Y-labeled antibody formats, derived from a single antimindin/RG-1 mAb, in a prostate tumor model. The IgG, diabody, single-chain variable domain (scFv), and novel miniantibody formats, composed of the human IgE-C(H)4 and a modified IgG1 hinge linked to scFv domains, were compared.
  
  
  METHODS
  Antibodies were first derivatized with the bifunctional chelator CHX-A''-diethylenetriamine pentaacetic acid and then bound to the radiometal to create radiolabeled immunoconjugates. Human LNCaP xenografts were grown in nude mice, and (111)In- or (86)Y-labeled antibodies were administered intravenously. Tissues were harvested at different times, and the level of antibody deposition was determined by measuring radioactivity. Whole-body small-animal PET of mice receiving (86)Y-labeled antibodies was performed at 6 time points and colocalized with simultaneous micro-CT imaging.
  
  
  RESULTS
  The biodistributions of (111)In and (86)Y antibodies were quite similar. The blood, tumor, kidney, and liver tissues contained varying levels of radioactivity. The antibody accumulation in the tumor correlated with molecular size. The IgG steadily increased with time to 24.1 percentage injected dose per gram (%ID/g) at 48 h. The miniantibody accumulated at a similar rate to reach a lower level (14.2 %ID/g) at 48 h but with a higher tumor-to-blood ratio than the IgG. Tumor accumulation of the diabody peaked at 3 h, reaching a much lower level (3.7 %ID/g). A combination of rapid clearance and lower relative affinity of the scFv precluded deposition in the tumor. Small-animal PET results correlated well with the biodistribution results, with similar tumor localization patterns.
  
  
  CONCLUSIONS
  The larger antibody formats (IgG and miniantibody) gave higher tumor uptake levels than did the smaller formats (diabody and scFv). These larger formats may be more suitable for radioimmunotherapy applications, evidenced by the preclinical efficacy previously shown by a report on the IgG format. The smaller formats were rapidly cleared from circulation, and the diabody, which accumulated in the tumor, may be more suitable for radiodiagnostic applications.
authors.affiliation: Cancer Research, Berlex Biosciences/Schering AG, Richmond, California, USA. doug.schneider@bayer.com
authors.forename: Douglas W
authors.initials: DW
authors.lastname: Schneider
authors.forename: Tara
authors.initials: T
authors.lastname: Heitner
authors.forename: Bruno
authors.initials: B
authors.lastname: Alicke
authors.forename: David R
authors.initials: DR
authors.lastname: Light
authors.forename: Kirk
authors.initials: K
authors.lastname: McLean
authors.forename: Noboru
authors.initials: N
authors.lastname: Satozawa
authors.forename: Gordon
authors.initials: G
authors.lastname: Parry
authors.forename: Jeongsoo
authors.initials: J
authors.lastname: Yoo
authors.forename: Jason S
authors.initials: JS
authors.lastname: Lewis
authors.forename: Renate
authors.initials: R
authors.lastname: Parry
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000493:pharmacokinetics*; D045744:Cell Line, Tumor; D002614:Chelating Agents / Q000737:chemistry; D006801:Humans; D007205:Indium Radioisotopes; D017879:Isothiocyanates / Q000737:chemistry; D008297:Male; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D004369:Pentetic Acid / Q000031:analogs & derivatives / Q000737:chemistry; D049268:Positron-Emission Tomography; D011471:Prostatic Neoplasms / Q000378:metabolism*; D019275:Radiopharmaceuticals / Q000737:chemistry / Q000493:pharmacokinetics*; D014018:Tissue Distribution; D014057:Tomography, X-Ray Computed; D014183:Transplantation, Heterologous; D015021:Yttrium Radioisotopes
chemical_list: D000911:Antibodies, Monoclonal; D002614:Chelating Agents; D007205:Indium Radioisotopes; D017879:Isothiocyanates; D019275:Radiopharmaceuticals; D015021:Yttrium Radioisotopes; C085678:N-(2-amino-3-(4-isothiocyanatophenyl)propyl)cyclohexane-1,2-diamine-N,N',N',N'',N''-pentaacetic acid; D004369:Pentetic Acid