Comparative Immunochemical Analysis of Wr^a and Wr^b Red Cell Antigens

Abstract

This paper describes immunoblotting and immunoprecipitation studies using monoclonal anti-Wr^a and anti-Wr^b antibodies to investigate the nature of the low- incidence blood group antigen, Wr^a and its high-incidence allelic antigen Wr^b. No membrane components were identified by the immunoblotting experiments. Immunoprecipitation studies confirmed that the Wr^b antigen involves both gly- cophorin A and band 3. The monoclonal anti-Wr ', BGU1-WR^a, failed to immu- noprecipitate these or any other red cell membrane component. The significance of these findings is discussed.

Diego Blood Group System

“…In GPA -defi cient cells the N -glycan on band 3 is of increased molecular weight and, despite normal quantities of band 3, anion transport is impaired (Sections 3.5.1 and 3.23), as a result of altered band 3 structure [106] . This is not applicable anti -Wr b either precipitated GPA [121,122] or GPA together with band 3 [123,124]. Red cells of band 3 knockout mice are defi cient in both band 3 and GPA, suggesting that band 3 plays a chaperone -like role and is essential for expression of GPA at the red cell surface [108] .…”

“…Attempts to identify any membrane component by immunoprecipitation with monoclonal anti -Wr a were unsuccessful [124] . Attempts to identify any membrane component by immunoprecipitation with monoclonal anti -Wr a were unsuccessful [124] .…”

Investigation of the Biochemical Relationship between the Blood Group Antigens Xg^a and CD99 (12E7 Antigen) on Red Cells

“…Immunoprécipitation of unlabelled or biotin-labelled red cells was carried out essentially according to the method described by Ring et al [12], Immune complexes were purified from the solubilized cells by incubating with protein-A-sepharose or rabbit anti-mouse-sepharose. For avidin-biotin immunoprécipitation, red cells were prelabelled with 0.7 mM N-hydroxysuccinimidobiotin (NHS-biotin) as described by Laing et al [13].…”

Investigation of the Biochemical Relationship between the Blood Group Antigens Xga and CD99 (12E7 Antigen) on Red Cells

“…For immunodetection of the glycoproteins, gels were electroblottcd and immunostained with antibodies as indicated, followed by incubation with peroxidase-labelled rabbit anti-human or rabbit antimouse Ig as described previously [l I]. lmm rmoyreci/~itution Immunoprecipitation of unlabelled or biotin-labelled red cells was carried out essentially according to the method described by Ring et al [12]. Immune complexes were purified from the solubilized cells by incubating with protein-A-sepharose or rabbit anti-mouse-sepharose.…”