Isolation and purification of Rh0(D) antigen of human erythrocyte membrane and its serological property.
To elucidate immunochemical properties of Rh-Hr antigens, influence of various solubilizers on the activity, and isolation of the antigens were investigated. As a very sensitive method to detect Rh-Hr antigens, the agglutination inhibition test was recommended in which diluted antisera (1:400-1:1,600), papainized red cells and microtitrate plates were used. Extraction of D antigen with Brij 35 and Triton X-100 was effective, but SDS was not successful. However, D activity of SDS solubilized stroma was recovered by adding reductants. The activity was slightly demonstrated at the preparation from D negative red cells. The influence of the reductants was discussed. SDS solubilized stroma was fractionated with gel filtrations and activated thiol Sepharose 4B. The final preparation with D activity was proved to correspond with Band 3 which was not stained with PAS reagent. Deoxycholic acid solubilized stroma was fractionated with gel filtrations and affinity chromatography, resulting in the preparation corresponding to Band 3 with potent Rh-Hr blood type activities. The activities were detected without reductants and the preparation from D negative red cells had no D activity. Rh-Hr blood type activities other than D were observed in the same fraction.
D000372:Agglutination Tests; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000032:analysis* / Q000276:immunology; D006801:Humans; D012204:Rh-Hr Blood-Group System / Q000276:immunology / Q000302:isolation & purification*; D013501:Surface-Active Agents / Q000494:pharmacology
D012204:Rh-Hr Blood-Group System; C026410:Rho(D) antigen; D013501:Surface-Active Agents