An anti-idiotopic antibody-based enzyme-linked immunosorbent assay far the quantification of the monoclonal anti-D BRAD-5

Objectives We aimed to develop an anti-idiotopic antibody-based enzyme-linked immunosorbent assay (ELISA) to quantify human monoclonal anti-D using BRAD-5 as a model system. Materials and Methods One of the anti-idiotopic antibodies 2E6 was used to capture BRAD-5 with the other anti-idiotopic antibody 3B1 biotinylated for detection. Results The assay developed can detect BRAD-5 at < 2 ng/ml. Assay interference caused by heterophilic antibodies in some human sera was removed by preincubation with bovine serum. The assay is reproducible with intra- and inter-assay coefficient of variation (CV) < 10%. Conclusion This ELISA should prove of benefit in developing a monoclonal anti-D for prophylactic use.