Examining technical aspects of the monoclonal antibody immobilisation of granulocyte antigen assay.
OBJECTIVE
This study was an attempt to improve detection and characterization of alloantibodies to neutrophil antigens, using the monoclonal antibody immobilisation of granulocyte antigens (MAIGA) assay.


METHODS
We explored the effect of different detergent concentrations, detergent class, stabilising additives, combinations of detergents, and use of a broad range of monoclonal antibodies on the results of the MAIGA assay.


RESULTS
Non-ionic detergents, Triton X and Nonidet, were confirmed as the most suitable for solubilisation of the recognised, neutrophil-associated alloantigens. Anionic and zwitterionic detergents did not assist the demonstration and characterisation of new neutrophil alloantigens in the MAIGA assay. Some reproducible positive MAIGA assay results were obtained for antisera 01, 08, anti-RED, anti-NB2, and anti-NB1, using particular monoclonal antibodies.


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 CD66b.
Minchinton
R M
RM
Australian Red Cross Blood Service, Queensland, Brisbane, Australia. 100240.1570@compuserve.com
Noonan
K
K
Johnson
T J
TJ
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis; D018845:CD11 Antigens / Q000032:analysis; D002793:Cholic Acids; D003840:Deoxycholic Acid; D003902:Detergents; D058851:GPI-Linked Proteins; D006098:Granulocytes; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D007518:Isoantibodies / Q000032:analysis*; D007519:Isoantigens / Q000032:analysis / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000032:analysis; D009504:Neutrophils / Q000737:chemistry; D017830:Octoxynol; D011092:Polyethylene Glycols; D011956:Receptors, Cell Surface; D017452:Receptors, IgG / Q000032:analysis; D012995:Solubility; D001613:beta 2-Microglobulin / Q000032:analysis
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D018845:CD11 Antigens; C407242:CD177 protein, human; D002793:Cholic Acids; D003902:Detergents; D058851:GPI-Linked Proteins; D007518:Isoantibodies; D007519:Isoantigens; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D017452:Receptors, IgG; D001613:beta 2-Microglobulin; D003840:Deoxycholic Acid; C024062:nonidet; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate