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Analysis of granulocyte‐reactive antibodies using an immunoassay based upon monoclonal‐antibody‐specific immobilization of granulocyte antigens
Author(s) -
Bux J.,
Kober B.,
Kiefel V.,
MuellerEckhardt C.
Publication year - 1993
Publication title -
transfusion medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.471
H-Index - 59
eISSN - 1365-3148
pISSN - 0958-7578
DOI - 10.1111/j.1365-3148.1993.tb00055.x
Subject(s) - granulocyte , monoclonal antibody , immunoassay , antibody , immunology , antigen , monoclonal , medicine , chemistry
Summary. To detect human granulocyte‐reactive antibodies, a glycoprotein‐specific enzyme immunoassay for platelet antibodies was adapted for the use of granulocytes as target cells. Peripheral blood granulocytes were simultaneously incubated with a monoclonal antibody (mAb) and the serum to be investigated. After solubilization, aliquots of the cell lysate were transferred to plastic tubes coated with goat anti‐mouse antibodies. Following immobilization of the trimolecular (mAb‐glycoprotein‐human antibody) complex it was detected by addition of enzyme‐labelled goat anti‐human antibodies using a luminescence technique. This assay allowed identification of different granulocyte‐reactive antibodies present in the same sample without the need for complicated absorption studies. Alloantibodies against HLA and the granulocyte‐specific NA antigens as well as isoantibodies against the Fc‐gamma‐receptor III (FcRIII) were detectable using mAb‐specific immobilization of granulocyte antigens (MAIGA). Binding of autoantibodies to the FcRIII and to the CD 11b/CD 18 complex could be shown.