TY - JOUR
T1 - Antibody affinity may influence antigenic modulation of the common acute lymphoblastic leukemia antigen in vitro
AU - Lebien, T. W.
AU - Boue, D. R.
AU - Bradley, J. G.
AU - Kersey, J. H.
PY - 1982
Y1 - 1982
N2 - 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 B-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 107 M-1 and 7.2 x 107 M-1 respectively, with NALM-6 cells expressing 1 x 105 to 2 x 105 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.
AB - 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 B-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 107 M-1 and 7.2 x 107 M-1 respectively, with NALM-6 cells expressing 1 x 105 to 2 x 105 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.
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M3 - Article
C2 - 6956632
AN - SCOPUS:0019921557
SN - 0022-1767
VL - 129
SP - 2287
EP - 2292
JO - Journal of Immunology
JF - Journal of Immunology
IS - 5
ER -