A glycoprotein of molecular weight 85,000 on human cells of B-lineage: detection with a family of monoclonal antibodies.
Immunization of BALB/c mice with glycoproteins purified from a detergent extract of human chronic lymphocytic leukemia (CLL) cells by affinity to Lens culinaris lectin led to the production of several monoclonal antibodies with similar reactivity. One of the antibodies, 50B4, was purified and the corresponding antigen was isolated from a B-lymphoblastoid cell line extract by affinity chromatography to the 50B4-IgG immunoadsorbent. Co-purification of the antigenic activities associated with five other monoclonal antibodies was achieved. Purified and radiolabelled 50B4 antigen could be specifically immunoprecipitated not only by 50B4 but also by the other five antibodies. SDS-PAGE analysis revealed that all antibodies precipitated the same component, a polypeptide chain of apparent mol. wt 85,000 under reducing conditions. Competitive-binding studies between the purified antibodies indicated the presence of two distinct epitopes on the antigen. The epitopes, each recognized by three different antibodies, were equally accessible on the cell surface of either a B-CLL (3 X 10(5) molecules/cell), a B-lymphoblastoid cell line (11 X 10(5) molecules/cell) or two acute lymphocytic leukemia (ALL) cell lines of pre-B phenotype (5 X 10(5) and 0.8 X 10(5) molecules/cell respectively). Although the antigens purified from the strongly positive ALL cell line gave a gel pattern identical to that of the B-lymphoblastoid cell line, the antigens purified from the B-CLL extract were resolved into two distinct glycosylated polypeptides of mol. wts 85,000 and 77,000 under reducing conditions. The distribution of the antigen(s) is not restricted to cells of the B-lineage as mature T-cells and a variety of non-hematopoietic cell types express both epitopes of the antigen(s).
Letarte
M
M
Iturbe
S
S
Quackenbush
E J
EJ
D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000951:Antigens, Neoplasm / Q000302:isolation & purification*; D000954:Antigens, Surface / Q000302:isolation & purification*; D001402:B-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D002846:Chromatography, Affinity; D000939:Epitopes / Q000032:analysis; D006023:Glycoproteins / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007163:Immunosorbent Techniques; D007945:Leukemia, Lymphoid / Q000276:immunology*; D008970:Molecular Weight; D013601:T-Lymphocytes / Q000276:immunology
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D000939:Epitopes; D006023:Glycoproteins