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The Use of the MAIEA Assay for Solving Serological Problems and Identifying Novel Blood Group Antigens
Author(s) -
Warke N.
Publication year - 2006
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.1365-3148.2006.00693_40.x
Subject(s) - epitope , monoclonal antibody , antigen , antibody , microbiology and biotechnology , serology , red blood cell , monoclonal , biology , chemistry , biochemistry , immunology
Background The monoclonal antibody‐specific immobilisation of erythrocyte antigens (MAIEA) assay was devised mainly for locating blood group antigens on specific red cell membrane proteins. The assay can also be used to map epitopes on protein molecules when used as a competitive binding assay, using monoclonal antibodies to different areas of a protein. The MAIEA assay has proved to be a very useful additional technique in the red cell reference laboratory when serological results are inconclusive and for assigning novel antigens to particular blood group systems. Methods The MAIEA assay detects trimolecular complexes formed by the reaction of an unknown human antibody and a known monoclonal antibody with a particular red cell protein. The trimolecular complex is captured on a microtitre plate coated with goat anti‐mouse immunoglobulin (Ig) and detected by the addition of peroxidase conjugated goat anti‐human Ig. A positive reaction indicates that the epitopes recognised by the human and monoclonal antibodies are present on the same membrane protein and are binding at different regions. A negative reaction indicates that the epitopes are either not on the same protein or the two antibodies are competing for the same or similar epitopes resulting in inhibition. Results During a 39 month period we carried out the MAIEA assay on 65 samples using the following monoclonal antibodies: CR1 (Knops) [54]; CD44 (Indian) [3]; AChE (Yt) [2]; DAF (Cromer) [3]; Kell [2]; Lutheran [1]. A suspected specificity was confirmed on 28 occasions: CR1 [21]; CD44 [3]; AChE [1]; DAF [1]; Kell [1]; Lutheran [1]. The Kell, Lutheran and two of the CD44 antibodies were to novel specificities which have since been defined by molecular analysis. Conclusions The MAIEA assay is a useful technique in the identification of unknown and novel specificities. Assigning a specificity to a particular blood group protein, followed by competitive binding assays, can indicate the region of the gene for molecular analysis. In addition to solving many difficult and complex serological problems, four novel antigens have been identified using the MAIEA assay over a 39 month period.