The binding of human alloantibodies to recombinant glycophorin A.
Chinese hamster ovary (CHO) cells were transfected with the wild-type, M allele of glycophorin A cDNA. The binding of human alloantibodies to recombinant glycophorin A was assessed with a modified hemagglutination-inhibition assay. Patient sera were incubated with acetone powders derived from CHO cells, and the adsorbed supernatants were tested in standard hemagglutination assays. Five M antibodies and one sample containing anti-En(a) bound to transfected CHO cells expressing glycophorin A but did not bind to untransfected CHO cells. Three N antibodies as well as 21 other alloantibodies (representing other major red cell blood group specificities) bound to neither CHO cell line. The M allele specificity of recombinant glycophorin A was further verified by the demonstration that a high-titer D alloantibody maintained the same titer of agglutination after incubation with recombinant glycophorin A. Transfected CHO cells thus express an M blood group antigen that appears to be serologically equivalent to that found on human red cells. A panel of cell lines expressing mutant glycophorin A molecules with defined variations in amino acid sequence and carbohydrate composition will be useful in studies of the fine specificity of human glycophorin alloantibodies. This approach may also provide an abundant source of artificial antigens for clinical use in blood group serology.
Blackall
D P
DP
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia.
Ugorski
M
M
Smith
M E
ME
Påhlsson
P
P
Spitalnik
S L
SL
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D006021:Glycophorins / Q000378:metabolism*; D006386:Hemagglutination Tests; D006801:Humans; D007518:Isoantibodies / Q000378:metabolism*; D008951:MNSs Blood-Group System / Q000276:immunology; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D012204:Rh-Hr Blood-Group System / Q000276:immunology; D014162:Transfection
D006021:Glycophorins; D007518:Isoantibodies; D008951:MNSs Blood-Group System; D011994:Recombinant Proteins; D012204:Rh-Hr Blood-Group System