Augmented binding of radiolabeled monoclonal antibodies to melanoma cells using specific antibody combinations.
Antigenic heterogeneity may limit effective cancer therapy using monoclonal antibodies (Mabs). To address this problem, combinations of two, three, or four 125I-labeled antimelanoma Mabs (NRML-05, P94, 96.5, and CL207) were incubated in vitro with three different melanoma cell lines (HS294t, A375SM, and DX3). Binding of the various Mab combinations was expressed as total cpm/10(5) cells and was compared to binding of each Mab alone. Saturating amounts (10 micrograms/ml) of two, three, or four Mabs bound to a significantly greater extent (P less than 0.05) than each individual Mab except for NRML-05. Combinations of three Mabs at a nonsaturating concentration (1.5 micrograms/ml) bound to a greater extent than single Mabs (P less than 0.05), depending on the cell line examined and the amount of antigen sites present for each Mab. Saturating or nonsaturating concentrations of unlabeled Mab 96.5 combined with 125I-labeled NRML-05 enhanced binding of the latter to HS294t by significantly modifying its affinity and by increasing the number of binding sites 3-fold. Modulation occurred only at 37 degrees C and was dependent upon protein synthesis. These data demonstrate that the effectiveness of various Mab combinations over single Mabs varies, depending on Mab concentration and the cell lines used. In addition, one Mab may significantly (P less than 0.05) enhance binding of another Mab to its antigen.
Mujoo
K
K
Department of Clinical Immunology and Biological Therapy, University of Texas M. D. Anderson Cancer Center, Houston 77030.
Rosenblum
M G
MG
Murray
J L
JL
D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000951:Antigens, Neoplasm; D006801:Humans; D007457:Iodine Radioisotopes; D007463:Iodobenzoates; D008545:Melanoma / Q000276:immunology / Q000378:metabolism*; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D007457:Iodine Radioisotopes; D007463:Iodobenzoates; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins