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An anti‐idiotopic antibody‐based enzyme‐linked immunosorbent assay for the quantification of the monoclonal anti‐D BRAD‐5
Author(s) -
Austin E. B.,
Smith L. C.,
Walker R. Y.
Publication year - 2001
Publication title -
vox sanguinis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.68
H-Index - 83
eISSN - 1423-0410
pISSN - 0042-9007
DOI - 10.1046/j.1423-0410.2001.00031.x
Subject(s) - monoclonal antibody , biotinylation , antibody , microbiology and biotechnology , chemistry , monoclonal , coefficient of variation , enzyme , chromatography , medicine , biology , immunology , biochemistry
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.