Coordinator's report on the flow cytometric analysis of the Rh antibodies.

One hundred and four IgG monoclonal antibodies with specificity within the Rh Blood Group System were evaluated by flow cytometry as part of the Third International Workshop. Standardisation of data to permit interlaboratory comparison of antibody binding was achieved by adherence to a standard red blood cell staining protocol, defined control cells and a standard FITC-labelled antibody. In addition, calibration standards were provided to permit the calculation of Molecules of Equivalent Soluble Fluorochrome (MESF) values from the mean channel fluorescence. For the majority of anti-D antibodies mean MESF values obtained with D positive cells were far higher than with the negative controls (D negative cells), with D variants having intermediate although very varied MESFs. In general MESF values obtained with non anti-D antibodies were less than for the anti-D antibodies although some of the anti-E antibodies were very strong. The highest MESF values were obtained with an anti-CD antibody.