title: 188Re-labeled anti-epidermal growth factor receptor humanized monoclonal antibody h-R3: labeling conditions, in vitro and in vivo stability.
abstract: The use of antibodies as targeting agents for the delivery of radioisotopes to tumors is an appealing concept that has received widespread attention since the advent of monoclonal antibody (mAb) technology. The present study describes the (188)Re-direct labeling of anti-epidermal growth factor receptor (EGF-R) humanized mAb h-R3; the analytical methods for quality control of radiopharmaceuticals such as instant thin layer chromatography-silica gel (ITLC-SG); the immunoreactivity and biological recognition of the target antigen assessment of the radiolabeled molecule using flow cytometry analysis; in vitro stability studies using saline 0.9% solution, cysteine, diethylenetriaminepentaacetic acid (DTPA), human serum and human serum albumin (HSA) 1% challenge; and the assessment of in vivo stability through biodistribution studies in normal Balb/c mice. No fragmentation of the reduced molecules was found using 2-ME as a reducing agent. Labeling efficiency was greater than 98.5 +/- 0.6% of rhenium-188 (188Re) bound to IgG1 after 5 h, as determined by paper chromatography in saline 0.9% solution. Radiocolloids determined by albumin impregnated ITLC was 1.04 +/- 0.07% in all cases. The biological activity measured by flow cytometry analysis showed an immunoreactivity fraction and the biological recognition of the target antigen overexpressed on H-125 human lung adenocarcinoma cell line greater than 87%. Challenge studies with cysteine, DTPA, human serum and HSA 1% demonstrated no evidence of transcomplexation of 188Re to DTPA or HSA and showed that 30% and 85% of the 188Re-radiolabeled was transcomplexed to human serum and to 100 mM cysteine after 24 h for human serum and 1 h incubation for cysteine at 37 masculine C, respectively. Biodistribution studies indicated no accumulation of the radiolabeled antibodies in normal organs.
authors.affiliation: Center of Molecular Immunology, Havana, Cuba.
authors.forename: N
authors.initials: N
authors.lastname: Iznaga-Escobar
authors.forename: I L
authors.initials: IL
authors.lastname: Ramirez
authors.forename: J C
authors.initials: JC
authors.lastname: Izquierdo
authors.forename: L
authors.initials: L
authors.lastname: Suárez
authors.forename: D
authors.initials: D
authors.lastname: Morales
authors.forename: R
authors.initials: R
authors.lastname: Pérez-Rodríguez
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D001666:Binding Sites, Antibody; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D004355:Drug Stability; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D011868:Radioisotopes / Q000032:analysis / Q000097:blood / Q000378:metabolism*; D012211:Rhenium / Q000032:analysis / Q000097:blood / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal; D011868:Radioisotopes; D012211:Rhenium; D066246:ErbB Receptors