title: Tumor-specific anti-epidermal growth factor receptor variant III monoclonal antibodies: use of the tyramine-cellobiose radioiodination method enhances cellular retention and uptake in tumor xenografts.
abstract: Amplification and rearrangement of the epidermal growth factor receptor (EGFR) gene are characteristics of many types of tumors. One class of EGFR mutations, EGFRvIII, is characterized by an in-frame deletion resulting in a truncated external domain of the receptor. EGFRvIII was first identified in a subset of gliomas and has since been found in some non-small cell lung carcinomas and breast carcinomas. mAbs specific for this variant form of EGFR but unreactive with the wild-type EGFR have been reported from our laboratory. This study further characterizes three of these antibodies. We determined, via radiolabeling techniques and immunofluorescence microscopy, that, after cell binding in vitro, the anti-EGFRvIII-specific mAbs internalize at 37 degrees C. Furthermore, subsequent to internalization, the antibodies were processed intracellularly, presumably by lysosomal degradation. We also examined the use of an alternative radiolabeling procedure that uses nonmetabolizable radio-iodinated tyramine cellobiose. Our results show that the tyramine cellobiose labeling method allows for greater tumor cell retention of radiolabel in vitro (76% for tyramine cellobiose and 27% for Iodo-Gen after 24 h). Paired-label biodistribution studies in athymic mice indicate that anti-EGFRvIII mAb L8A4 localizes specifically to EGFRvIII-expressing tumor xenografts with a maximum of 34.3 +/- 7.6% injected dose/g when labeled using tyramine cellobiose compared with a maximum of 14.9 +/- 4.3% injected dose/g using Iodo-Gen; similar results were obtained with mAb H10. These results suggest that the anti-EGFRvIII mAbs may serve as potential carriers for radioconjugate- and immunotoxin-based therapies for tumors expressing EGFRvIII.
authors.affiliation: Department of Pathology, Duke University Medical Center, Durham, North Carolina 27710, USA.
authors.forename: C J
authors.initials: CJ
authors.lastname: Reist
authors.forename: G E
authors.initials: GE
authors.lastname: Archer
authors.forename: S N
authors.initials: SN
authors.lastname: Kurpad
authors.forename: C J
authors.initials: CJ
authors.lastname: Wikstrand
authors.forename: G
authors.initials: G
authors.lastname: Vaidyanathan
authors.forename: M C
authors.initials: MC
authors.lastname: Willingham
authors.forename: D K
authors.initials: DK
authors.lastname: Moscatello
authors.forename: A J
authors.initials: AJ
authors.lastname: Wong
authors.forename: D D
authors.initials: DD
authors.lastname: Bigner
authors.forename: M R
authors.initials: MR
authors.lastname: Zalutsky
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000493:pharmacokinetics* / Q000627:therapeutic use; D002475:Cellobiose; D004337:Drug Carriers; D066246:ErbB Receptors / Q000032:analysis / Q000276:immunology*; D007457:Iodine Radioisotopes / Q000493:pharmacokinetics*; D007553:Isotope Labeling / Q000379:methods*; D051379:Mice; D008856:Microscopy, Fluorescence; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000378:metabolism* / Q000628:therapy; D011829:Radiation Dosage; D014018:Tissue Distribution; D014183:Transplantation, Heterologous; D014439:Tyramine
chemical_list: D000911:Antibodies, Monoclonal; D004337:Drug Carriers; D007457:Iodine Radioisotopes; D002475:Cellobiose; D066246:ErbB Receptors; D014439:Tyramine