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Use of the DAF Assay to Assess the Functional Properties of Polyclonal and Monoclonal Rh D Antibodies
Author(s) -
Ducrot T.,
Beliard R.,
Glacet A.,
Klein P.,
Harbonnier S.,
Benmostefa N.,
Bourel D.
Publication year - 1996
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.1423-0410.1996.7110030.x
Subject(s) - antibody dependent cell mediated cytotoxicity , polyclonal antibodies , monoclonal antibody , phagocytosis , monoclonal , antibody , microbiology and biotechnology , peripheral blood mononuclear cell , chemistry , lysis , in vitro , cytotoxicity , immunology , biology , biochemistry
The mechanism whereby passive Rh (D) immunoglobulins suppress the feto‐maternal alloimmunization is still unclear. New in vitro tests are needed to better characterize the functional properties of polyclonal anti‐Ds. The DAF assay was developped to monitor the antibody‐dependent cell‐mediated cytotoxicity (ADCC) and the phagocytosis of anti‐Rh (D)‐sensitized RBCs by effector cells. The principle of this test is based on the oxydization of the 2,7‐diaminofluorene (DAF) by the pseudoperoxidase activity of free hemoglobin. The reaction is proportional to the hemoglobin concentration. This test was performed to determine and emphasize the efficacy of different polyclonal anti‐D immunoglobulin preparations to mediate lysis and phagocytosis of sensitized RBCs by human peripheral mononuclear cells. The functional properties of different human RhD monoclonal antibodies were also analyzed and compared. The test was found to be convenient to perform and allowed the avoidance of radioactive labelling of RBCs for ADCC studies. It is mainly useful for the direct quantitation of phagocytosis.