Identification and characterization of the anti-Duv^a+ antibody by the techniques of MAIPA and Western blot.

(A) The reactivity of the serum Duv was tested in MAIPA with the mother's platelets, 4 panel platelets, and the father's platelets by using the Moab Pl1-64 to capture the GP IIb-IIIa complex. Paternal autoantibodies were searched for by direct incubation of the platelets with the Moabs (Dir). (B) Immunoblot of SDS-soluble platelet proteins subjected to SDS–polyacrylamide gel electrophoresis under nonreduced conditions and transferred to nitrocellulose membrane. The maternal serum Duv was tested with father's platelets (lane 1), mother's platelets (lane 2), and a control donor (lane 3). An anti–HPA-1a serum (anti–GP IIIa) was used as positive control (lane 4). Human IgGs were revealed by using an antihuman IgG conjugated to the peroxidase associated to the chemiluminescence detection technique. The maternal serum Duv bound specifically to a protein migrating in the position of GP IIIa of the father's platelets.