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


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%.


CONCLUSIONS
This ELISA should prove of benefit in developing a monoclonal anti-D for prophylactic use.
Austin
E B
EB
Manchester Blood Centre, Plymouth Grove, Manchester M13 9LL, UK. eric.austin@nbs.nhs.uk
Smith
L C
LC
Walker
R Y
RY
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology*; D000910:Antibodies, Heterophile / Q000097:blood; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000276:immunology; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007130:Immunoglobulin Idiotypes / Q000276:immunology; D007518:Isoantibodies / Q000032:analysis* / Q000276:immunology; D051379:Mice; D015203:Reproducibility of Results; D012203:Rh Isoimmunization / Q000175:diagnosis; D018029:Rho(D) Immune Globulin; D012680:Sensitivity and Specificity; D013045:Species Specificity
D000888:Antibodies, Anti-Idiotypic; D000910:Antibodies, Heterophile; D000911:Antibodies, Monoclonal; D007130:Immunoglobulin Idiotypes; D007518:Isoantibodies; C061961:RHO(D) antibody; D018029:Rho(D) Immune Globulin