The monoclonal antibody‐specific immobilization of erythrocyte antigens assay (MAIEA) in the investigation of human red‐cell antigens and their associated membrane proteins

Abstract

The monoclonal antibody-specific immobilization of erythrocyte antigens (MAIEA) technique is an immunoassay devised primarily for locating blood group antigens on specific red-cell membrane proteins. The assay involves the incubation of intact red cells with two antibodies, one human alloantibody, the other a nonhuman antibody, usually a rodent monoclonal antibody, but polyclonal antibodies of rabbit origin have been utilized. For a positive result, both antibodies must bind to the same membrane protein. The red cells are lysed, the membrane solubilized and the trimolecular complex of two antibodies and membrane protein is captured in a well coated with goat antirodent (or rabbit) immunoglobulin. The immobilized complex is then detected by the use of peroxidase-conjugated goat antihuman (or rodent) immunoglobulin. Negative results, due to mutual blocking between the human and animal antibodies when their epitopes are close together on the same molecule, have permitted a degree of localization of epitopes on some proteins. This has been most effective in the mapping of Cromer blood group system antigens on the complement control protein domains of decay-accelerating factor (DAF, CD55), but has also proved informative in the clustering of antigens on the Lutheran and Kell glycoproteins. MAIEA is an effective tool for the identification of antibodies to Knops-system antigens on complement receptor 1 (CR1, CD35) in immunohaematology reference laboratories. These antibodies are clinically unimportant, but must be identified before they can be ignored for transfusion purposes.

Recombinant blood group proteins facilitate the detection of alloantibodies to high‐prevalence antigens and reveal underlying antibodies: results of an international study

“…† DAF‐MAIEA = MAIEA test for detection of Cromer antibodies on the decay‐accelerating factor (DAF, CD55) …”

“…In selected cases, the IBGRL in Bristol used the monoclonal antibody–specific immobilization of erythrocyte antigens (MAIEA) technique for detection of antibodies to Knops system antigens on Complement Receptor 1 (CR1, CD35) and to Cromer antigens on decay‐accelerating factor (CD55). This immunoassay technique is designed primarily to locate blood group antigens on specific RBC membrane proteins and does not allow for precise localization of antigens on proteins …”

“…This immunoassay technique is designed primarily to locate blood group antigens on specific RBC membrane proteins and does not allow for precise localization of antigens on proteins. 14…”

Serological tools for investigating immunohaematology problems

“…Monoclonal antibodies specific for the suspected red cell membrane protein are selected and a positive result indicates the unknown human antibody has bound to the same protein as the monoclonal antibody/antibodies of known specificity [7]. This technique is particularly effective for identifying CR1 (Knops)-related specificities and for helping to assign novel specificities to particular blood group systems.…”

Immunohematology of the Kell and KX Blood Group Systems

“…Competitive antibody-binding assays have been used in an attempt to identify clusters of antigen activity [3,17]. Very little correlation between spatial positioning as determined by competitive binding and linear positioning is obvious.…”