Use of immobilized platelet membrane glycoproteins for the detection of platelet-specific alloantibodies in solid-phase ELISA.
Platelet membrane glycoproteins were isolated from intact platelets by detergent-phase extraction, fixed to the wells of microtiter trays and used as targets for the detection of platelet-reactive alloantibodies by enzymelinked immunospecific assay (ELISA). The final preparations contained 0.4% of total platelet protein. Antibodies reactive with antigens PlA1, PlA2, Baka, Pena and HLA-A2 were specifically detected at dilutions ranging form 1:640 to 1:1.600. Under the conditions utilized, the ELISA was more sensitive than assays involving 51Cr, radiolabeled monoclonal anti-IgG binding, and indirect immunofluorescence testing by one order of magnitude or greater. When platelets were pretreated with chloroquine to remove class I HLA antigens prior to detergent-phase extraction, reactions with HLA-specific antibodies were lost, but reactions with platelet-specific alloantibodies were retained. This approach offers a simple, sensitive and rapid method to detect and identify platelet-specific alloantibodies in sera containing HLA-reactive alloantibodies.
Collins
J
J
Blood Center of Southeastern Wisconsin, Medical College of Wisconsin, Milwaukee.
Aster
R H
RH
D000906:Antibodies / Q000032:analysis; D001792:Blood Platelets / Q000276:immunology*; D002738:Chloroquine; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D005069:Evaluation Studies as Topic; D005455:Fluorescent Antibody Technique; D006680:HLA Antigens / Q000276:immunology; D006801:Humans; D007518:Isoantibodies / Q000032:analysis*; D010980:Platelet Membrane Glycoproteins
D000906:Antibodies; D006680:HLA Antigens; D007518:Isoantibodies; D010980:Platelet Membrane Glycoproteins; D002738:Chloroquine