title: In vivo targeting of malignant melanoma by 125Iodine- and 99mTechnetium-labeled single-chain Fv fragments against high molecular weight melanoma-associated antigen.
abstract: Monoclonal antibodies (MAbs) against high-molecular-weight melanoma-associated antigen (HMW-MAA) have been used in vivo to target melanoma. More recently, single chain Fv (scFv) antibody fragments against HMW-MAA have been described that may improve melanoma targeting. However, there have been few in vivo studies with antimelanoma scFvs because these have proved difficult to label with isotopes (e.g., 99mTc) suitable for imaging. We have generated a series of scFvs against HMW-MAA by chain shuffling and antibody phage selection on melanoma cells. In preliminary experiments we identified one scFv (RAFT3) as suitable for in vivo melanoma targeting. Direct radiolabeling of RAFT3 scFv with 99mTc was simple, yielding a radiochemical purity of >90%. The label remained stable for 24 h in vitro. 125I- and 99mTc-labeled RAFT3 scFv were tested in a nude mouse xenograft model for human melanoma and were compared with the parent MAb LHM2 and its F(ab')2 fragment versus nonmelanoma-specific MAb and scFv. RAFT3 scFv accumulated specifically in the tumor and showed greater tumor specificity compared with LHM2 with faster pharmacokinetics (t(1/2)alpha, 8 min; t(1/2)beta, 189 min; and t(1/2)alpha, 37 min; t(1/2)beta, 384 min, respectively) and reduced background in liver, lung, and spleen. Nonspecific accumulation of 99mTc-labeled RAFT3 scFv in the kidney was high but tumor:normal tissue ratios were better compared with 125I-labeled RAFT3 scFv and LHM2 F(ab')2. Overall, tumor-targeting efficiency at equivalent time points was scFv > IgG > F(ab')2 in good agreement with previously described scFvs engineered for 99mTc labeling. We discuss the potential use of RAFT3 scFv for imaging and therapy of metastatic melanoma.
authors.affiliation: The Restoration of Appearance and Function Trust Institute of Plastic Surgery, Mount Vernon Hospital, Northwood, Middlesex, United Kingdom.
authors.forename: N
authors.initials: N
authors.lastname: Kang
authors.forename: S
authors.initials: S
authors.lastname: Hamilton
authors.forename: J
authors.initials: J
authors.lastname: Odili
authors.forename: G
authors.initials: G
authors.lastname: Wilson
authors.forename: J
authors.initials: J
authors.lastname: Kupsch
mesh_terms: D000818:Animals; D001435:Bacteriophages / Q000378:metabolism; D002852:Chromatography, Ion Exchange; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007128:Immunoglobulin Fragments / Q000494:pharmacology* / Q000627:therapeutic use; D007074:Immunoglobulin G / Q000378:metabolism; D007457:Iodine Radioisotopes / Q000627:therapeutic use*; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D008545:Melanoma / Q000188:drug therapy* / Q000276:immunology*; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D013154:Spleen / Q000378:metabolism; D013667:Technetium / Q000627:therapeutic use*; D013997:Time Factors; D014407:Tumor Cells, Cultured
chemical_list: D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; D007457:Iodine Radioisotopes; D013667:Technetium