Production of heavy-chain class-switch variants of human monoclonal antibody by recombinant DNA technology.
We have previously established a human-mouse heterohybridoma (H6-3C4), which produced a human sperm-immobilizing antibody (mu, lambda of human type). The human rearranged immunoglobulin mu-chain and lambda-chain genes were cloned from the hybridoma H6-3C4. The cloned V region of the heavy chain (VH) gene was ligated to human immunoglobulin gamma 1-heavy chain constant region (C gamma 1) genes. This resulted in the heavy-chain class-switch from mu-chain to gamma 1-chain of H6-3C4 antibody. The class-switched heavy-chain gene as well as the cloned lambda-chain gene were introduced into mouse myeloma cell line X63Ag8.653 by protoplast fusion and electroporation. The stable transformants produced the human IgG monoclonal antibody, which fully retained specificity to human sperm cells and sperm-immobilizing activity.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004591:Electrophoresis, Polyacrylamide Gel; D005803:Genes, Immunoglobulin; D006801:Humans; D007127:Immunoglobulin Constant Regions / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007146:Immunoglobulin lambda-Chains / Q000235:genetics; D007148:Immunoglobulin mu-Chains / Q000235:genetics; D007136:Immunoglobulins / Q000096:biosynthesis; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D013081:Sperm Motility; D013094:Spermatozoa / Q000276:immunology; D014162:Transfection
D000911:Antibodies, Monoclonal; D004274:DNA, Recombinant; D007127:Immunoglobulin Constant Regions; D007143:Immunoglobulin Heavy Chains; D007146:Immunoglobulin lambda-Chains; D007148:Immunoglobulin mu-Chains; D007136:Immunoglobulins