Development and properties of a monoclonal antibody specific for human ecto-5'-nucleotidase.
After immunization with purified human placental ecto-5'-nucleotidase and fusion, 18 different hybrids were obtained which produce an antibody inhibitory for enzyme activity. These antibodies show complete cross-reactivity with the enzyme in a membrane-bound form or on the surface of intact cells. After cloning and ascites production, the antibody of one clone ( IFH - 5N1 ) was studied in greater detail. The IFH - 5N1 antibody is of the IgG 1 subclass. Optimal enzyme inhibition is 90% for purified 5'-nucleotidase and 80% for the enzyme on lymphoblasts. The specificity of this antibody is further demonstrated by enzyme inhibition assays and fluorescence labeling using various 5'-nucleotidase-positive and -negative human cells such as peripheral blood lymphocytes, leukemic cells, lymphoblastoid B- and T-cell-lines and fibroblasts. The antibody should provide a useful tool for the diagnosis of certain forms of acute leukemias and for the study of normal human lymphocyte subpopulations.
D015720:5'-Nucleotidase; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001665:Binding Sites; D003429:Cross Reactions; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007945:Leukemia, Lymphoid / Q000201:enzymology; D008214:Lymphocytes / Q000201:enzymology; D051379:Mice; D009708:Nucleotidases / Q000276:immunology*
D000911:Antibodies, Monoclonal; D009708:Nucleotidases; D015720:5'-Nucleotidase