Monoclonal antibodies recognizing epitopes on the extracellular face and intracellular N-terminus of the human erythrocyte anion transporter (band 3) and their application to the analysis of South East Asian ovalocytes.
This report describes the production and characterization of 13 rodent monoclonal antibodies to the human erythrocyte anion transport protein AE1 (syn. band 3). Eleven antibodies (4 murine and 7 rat) recognize epitopes dependent on the integrity of the third extracellular loop of the protein. Two antibodies (1 murine and 1 rat) recognize epitopes on the N-terminal cytoplasmic domain. Quantitative binding studies using radioiodinated IgG and Fab fragments of antibodies to extracellular epitopes on AE1 ranged from 77,000 to 313,000 (IgG) and from 241,000 to 772,000 (Fab) molecules bound at saturation. The results indicate that the epitopes recognized by different antibodies vary in their accessibility and suggest that there is heterogeneity in the organization of individual AE1 molecules in the red blood cell membrane. Quantitative binding studies on South East Asian ovalocytes using several antibodies to AE1 and an anti-Wrb show a marked reduction in the number of antibody molecules bound at saturation. These results are consistent with the existence of highly cooperative interactions between transmembrane domains of AE1 in normal erythrocytes and the disruption of these interactions in the variant AE1 found in South East Asian ovalocytes.
Smythe
J S
JS
International Blood Group Reference Laboratory, Bristol, UK.
Spring
F A
FA
Gardner
B
B
Parsons
S F
SF
Judson
P A
PA
Anstee
D J
DJ
D000818:Animals; D001457:Anion Exchange Protein 1, Erythrocyte / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D004612:Elliptocytosis, Hereditary / Q000276:immunology*; D018604:Epitope Mapping; D004913:Erythrocytes, Abnormal / Q000276:immunology*; D005110:Extracellular Space; D006388:Hemagglutinins; D006801:Humans; D051379:Mice; D051381:Rats
D001457:Anion Exchange Protein 1, Erythrocyte; D000911:Antibodies, Monoclonal; D006388:Hemagglutinins