Human monoclonal antibody to tumor-associated ganglioside GD2.

Abstract

Human IgM kappa antibody to a membrane antigen of human tumors of neuroectodermal origin (melanoma, glioma and neuroblastoma) has been detected in the spent culture fluid of an Epstein-Barr virus (EBV)-transformed human B-lymphoblastoid cell line, L72. The chemical nature of the antigen was identified as ganglioside GD2. The antibody was purified by precipitation of L72 culture fluid with ammonia sulfate and hypotonic buffer followed by ultracentrifugation and Sephacryl S-300 super gel filtration. Approximately 27 mg of pure human IgM was obtained from 101 of spent medium. Total IgM and antibody activity recovery efficiency was 60% and 75%, respectively. The monoclonal character of the immunoglobulin produced by the L72 cell line was determined by agarose isoelectric focusing and immunofixation techniques. 1 mg of the purified IgM possessed an antibody titer endpoint to a GD2-positive melanoma cell line of 1:10,000 as assayed by immune adherence and 1:100 titer by complement-dependent cytotoxicity in vitro. The effect of pure anti-GD2 on suppression of melanoma growth in vivo was tested using a nude mouse model. Three-week-old CD-1 nude mice bearing 2-3 mm M14-A subcutaneous melanoma nodules were treated intraperitoneally with anti-GD2 and rabbit complement. Tumor growth was retarded for 25 days when compared to that of control mice receiving non-specific human IgM and complement. On Day 15, treated tumors were 80% smaller than control tumors. These result indicated that the pure human monoclonal antibody to GD2 may have potential for cancer therapy.

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@article{Katano1984HumanMA, title={Human monoclonal antibody to tumor-associated ganglioside GD2.}, author={Mitsuru Katano and Romaine E. Saxton and Reiko F . Irie}, journal={Journal of clinical & laboratory immunology}, year={1984}, volume={15 3}, pages={119-26} }