Antibody affinity may influence antigenic modulation of the common acute lymphoblastic leukemia antigen in vitro.
We have produced a new monoclonal antibody designated BA-3. An extensive side-by-side serologic comparison of BA-3 with the anti-common acute lymphoblastic leukemia antigen (CALLA) monoclonal antibody J-5 was undertaken. Cells examined included established leukemic cell lines, malignant cells from patients with newly diagnosed leukemia/lymphoma, and normal hematopoietic tissues. In all experiments the cellular distribution of the antigens recognized by BA-3 and J-5 were identical when analyzed by immunofluorescent microscopy and the FACS. Iodination of NALM-6 cells, followed by radioimmunoprecipitation and SDS-PAGE, indicated that BA-3 (like J-5) precipitated a glycoprotein of approximately 100,000 daltons. Competitive binding studies using 125I-labeled BA-3 indicated that BA-3 and J-5 were binding to closely associated (if not identical) epitopes on CALLA. Calculation of the equilibrium constant (K value) for BA-3 and J-5, and the approximate number of CALLA molecules per cell, was graphically determined using Scatchard plots. BA-3 and J-5 were shown to have K values of approximately 2.7 x 10(7) M-1 and 7.2 x 10(7) M-1, respectively, with NALM-6 cells expressing 1 x 10(5) to 2 x 10(5) CALLA molecules per cell. Additional studies with BA-3 failed to demonstrate significant antigenic modulation of CALLA in vitro using fresh leukemic cells and leukemic cell lines. Thus, we suggest that antibody affinity may be a significant factor influencing antigenic modulation of CALLA in vitro.
D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000915:Antibody Affinity; D000951:Antigens, Neoplasm / Q000032:analysis / Q000276:immunology*; D001666:Binding Sites, Antibody; D002460:Cell Line; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D006801:Humans; D007945:Leukemia, Lymphoid / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm