Recombinant human-mouse chimeric monoclonal antibody specific for common acute lymphocytic leukemia antigen.
A human-mouse chimeric antibody constructed in the present study was specific for a human tumor-associated antigen, common acute lymphocytic leukemia antigen. The antibody consisted of human heavy and light chain constant domains (gamma 1 and kappa type) and mouse heavy and light chain variable domains, which were derived from human plasma cell leukemia line (ARH77) and mouse hybridoma cells (NL-1) specific for common acute lymphocytic leukemia antigen, respectively. The artificially fused immunoglobulin molecules were produced in mouse myeloma cells, X63Ag8.653 which were transformed with the chimeric heavy and light chain genes formed by joining the corresponding gene segments in vitro at the J-C introns. The human heavy chain enhancer element was ligated tot he chimeric heavy and light chain genes, and this enhancer appeared to be obligatory for the efficient production of the chimeric antibody molecules. The stably transformed cells secreted the chimeric antibody, which specifically bound a common acute lymphocytic leukemia antigen expressing cell line. The amount of the chimeric antibody produced (10-30 micrograms/ml in the serum-free medium) was comparable to that made by murine hybridoma line, NL-1. The molecular weight of the chimeric heavy chain molecules was reduced from 54,000 to 50,000 upon treatment with tunicamycin, suggesting that the peptide was normally glycosylated in the transformants. The chimeric antibody exhibited complement-dependent cytotoxicity, in which glycosylation is thought to be indispensable. The antibody also mediated antibody-dependent cell-mediated cytotoxicity to the human target cells. The antibody-dependent cell-mediated cytotoxicity activity of the chimeric antibody was twice that of the murine NL-1 monoclonal antibody when human peripheral blood mononuclear cells were used as effectors.
D000818:Animals; D000911:Antibodies, Monoclonal; D000920:Antibody-Dependent Cell Cytotoxicity; D000951:Antigens, Neoplasm / Q000276:immunology*; D002460:Cell Line; D005453:Fluorescence; D006031:Glycosylation; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000276:immunology; D051379:Mice; D008970:Molecular Weight; D015260:Neprilysin; D011994:Recombinant Proteins / Q000276:immunology*; D014415:Tunicamycin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D007143:Immunoglobulin Heavy Chains; D011994:Recombinant Proteins; D014415:Tunicamycin; D015260:Neprilysin