Detection of antibodies reacting with the antithetical duffy blood group antigens Fy(a) and Fy(b) using recombinant fusion proteins containing the duffy extracellular domain.
Detecting blood group-specific antibodies in patient sera is essential to the management of blood transfusions or pregnancies. We produced the antithetical forms of the 65 amino acid extracellular domain (ECD) of the Duffy (Fy) blood group protein fused to glutathione sulfotransferase (GST): GST-Fy(a); and GST-Fy(b), differing only in Gly or Asp at position 44, respectively. The purified recombinant proteins were recognized more effectively by reference polyclonal or monoclonal antibodies than the antithetical Fy specificity by either ELISA or immunoblotting. Combined immunoblot and ELISA tests performed at 1:200 dilutions of sera using the recombinant proteins gave results in agreement with undiluted sera and agglutination for 17/19 alloimmunized patients. At 1:200, agglutination detected anti-Fy(a) or anti-Fy(b) in only three of 12 samples that were positive by ELISA. Recombinant ECD-Fy proteins are suitable and sensitive reagents for the detection of anti-Fy that use technology amenable to automation and/or miniaturization and avoid the need for intact red cells.
Sheffield
William P
WP
Canadian Blood Services, Research and Development, Hamilton and Toronto, Canada.
Bhakta
Varsha
V
Branch
Donald R
DR
Denomme
Gregory A
GA
D000372:Agglutination Tests; D000596:Amino Acids / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000097:blood*; D000937:Antigen-Antibody Reactions; D004375:Duffy Blood-Group System / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005982:Glutathione Transferase / Q000235:genetics / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology*; D012680:Sensitivity and Specificity
C473745:ACKR1 protein, human; D000596:Amino Acids; D000911:Antibodies, Monoclonal; D004375:Duffy Blood-Group System; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase