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Use of Immobilized Platelet Membrane Glycoproteins for the Detection of Platelet‐Specific Alloantibodies in Solid‐Phase ELISA 1
Author(s) -
Collins Janice,
Aster Richard H.
Publication year - 1987
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 - 10.1111/j.1423-0410.1987.tb04941.x
Subject(s) - platelet , antigen , chemistry , monoclonal antibody , platelet membrane glycoprotein , glycoprotein , antibody , microbiology and biotechnology , human leukocyte antigen , isoantibodies , serial dilution , chromatography , immunology , biochemistry , biology , medicine , pathology , alternative medicine
. 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 enzyme‐linked immunospecific assay (ELISA). The final preparations contained 0.4% of total platelet protein. Antibodies reactive with antigens P1 A1 , P1 A2 , Bak a , Pen a 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 51 Cr, 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.