Production of human monoclonal IgG antibodies against Rhesus (D) antigen.

Publisher Website
Google Scholar
Abstract
An Epstein-Barr virus (EBV)-transformed human B cell line (LB4r) producing anti-Rhesus [Rho(D) antigen] antibody was fused with a non-Ig-producing mouse-human heteromyeloma (SHM-D33) and selected in hypoxanthine/aminopterin/thymidine medium containing 0.5 .mu.M ouabain. Surviving hybrids secreted specific anti-Rho(D) antibody were cloned by limiting dilution. Two clones (D4-B2 and E10-C1) producing high levels (12 and 20 .mu.g/ml per 106 cells per 24 h, respectively) of monospecific antibody (IgG3, .lambda. chain) were selected for expansion and further characterization. Compared to the parental cell line (LB4r), these hybridoma cell lines presented several advantages: antibody production was increased 10-fold, cloning efficiency was improved, and the EBV genome was not retained. Antibody production has been stable for > 8 mo. These human monoclonal anti-Rho(D) antibodies have demonstrated utility in routine blood-group typing. They may also prove useful in the biochemical and genetic characterization of the Rh antigen system. Most important, they offer a source of Rh-immune globulin for the prevention of Rh immunization and alloimmune hemolytic disease of the newborn.