EXAMINING TECHNICAL ASPECTS OF THE MONOCLONAL-ANTIBODY IMMOBILIZATIONOF GRANULOCYTE ANTIGEN-ASSAY

Background and Objectives: This study was an attempt to improve detect ion and characterization of alloantibodies to neutrophil antigens, usi ng the monoclonal antibody immobilisation of granulocyte antigens (MAI GA) assay. Materials and Methods: We explored the effect of different detergent concentrations, detergent class, stabilising additives, comb inations of detergents, and use of a broad range of monoclonal antibod ies on the results of the MAIGA assay. Results: Non-ionic detergents, Triton X and Nonidet, were confirmed as the most suitable for solubili sation of the recognised, neutrophil-associated alloantigens. Anionic and zwitterionic detergents did not assist the demonstration and chara cterisation of new neutrophil alloantigens in the MAIGA assay. Some re producible positive MAIGA assay results were obtained for antisera 01, 08, anti-RED, anti-NB2, and anti-NB1, using particular monoclonal ant ibodies. Conclusions: Although we are reluctant to suggest association of the targeted antigens with the particular glycoproteins identified by the capture monoclonal antibodies, NB1 glycoprotein may be closely associated, before or after antibody binding, with CD46, CD11b, CD50, and/or CD 66b.