A new photometric method for the quantitation of Fc receptors for murine IgG1 on human monocytes and cell lines.
We have previously shown a polymorphism of human Fc receptors for mouse IgG1 using an EA rosette technique in which human erythrocytes sensitized with a murine IgG1 monoclonal antibody against glycophorin A acted as indicator cells. We now describe a method to quantitate this EA rosetting using the pseudoperoxidase activity present in erythrocytes. This photometric assay allows the sensitive quantitative determination of Fc receptor expression on human monocytes and cell lines. Not only the human Fc receptor for murine IgG1 can be studied in this way, but the method can also be applied to other Fc receptors. An important factor in this type of rosette assay appears to be the amount of negative charge present on the surface of the indicator erythrocytes. Using alcian blue as a probe, we found that this negative charge is higher on human erythrocytes than on sheep erythrocytes, which may contribute to a better signal-to-noise ratio. The method described facilitates the characterization of Fc receptors and permits the rapid screening of monoclonal anti-Fc receptor antibodies.
D000042:Absorption; D000818:Animals; D000906:Antibodies / Q000276:immunology; D002460:Cell Line; D004912:Erythrocytes / Q000201:enzymology / Q000276:immunology; D006454:Hemoglobins / Q000032:analysis / Q000502:physiology; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism*; D051379:Mice; D009000:Monocytes / Q000648:ultrastructure*; D010544:Peroxidases / Q000378:metabolism; D010783:Photometry / Q000379:methods*; D011961:Receptors, Fc / Q000032:analysis* / Q000276:immunology; D012397:Rosette Formation / Q000379:methods
D000906:Antibodies; D006454:Hemoglobins; D007074:Immunoglobulin G; D011961:Receptors, Fc; D010544:Peroxidases