Identification and quantitation of the protein content of blood group antibody eluates prepared by the rapid acid-stromal method. Use in committed erythroid progenitor cell assays.
Anti-D sensitized red cells were converted to stroma, either by digitonin or hypotonic lysis, and eluates prepared by the modified rapid acid-stromal method. SDS-PAGE and both rocket- and crossed-immunoelectrophoresis showed that, of the total protein content of the eluates, 11 to 25 percent was IgG, 18 to 45 percent was globin, and 34 to 71 percent was red cell membrane protein. Eluates were free of albumin, transferrin, and other non-IgG serum proteins; they did not contain proteolytic activity, and eluates were noninhibitory to the growth of committed erythroid progenitors. The rapid acid-stromal method produced high-titered blood group antibody eluates free of contaminating immune or nonimmune activity which were suitable for assay of blood group antigen expression by hemopoietic stem cells and committed progenitors.
D001798:Blood Proteins / Q000032:analysis*; D003378:Counterimmunoelectrophoresis; D004072:Digitonin / Q000494:pharmacology; D004912:Erythrocytes / Q000032:analysis* / Q000187:drug effects; D005914:Globins / Q000032:analysis; D006412:Hematopoietic Stem Cells / Q000032:analysis*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D007518:Isoantibodies / Q000276:immunology; D008565:Membrane Proteins / Q000032:analysis
D001798:Blood Proteins; D007074:Immunoglobulin G; D007518:Isoantibodies; D008565:Membrane Proteins; D005914:Globins; D004072:Digitonin