A quantitative analysis of cell surface glycosphingolipid with a fluorescence activated cell sorter

The fluorescence activated cell sorter (FACS) was used with an indirect membrane immunofluorescence technique to detect antibody against the Forssman antigen, a glycosphingolipid. Sheep erythrocytes, which contain Forssman antigen as a major membrane glycosphingolipid, were used as the target antigen. Detection of the anti-Forssman antibody on the sheep erythrocytes was done with specific fluorescein-conjugated second antibody and analyzed on a FACS. Compared to other available methods, analysis with the FACS was simple, sensitive, reproducible and quantitative. More than 250 pg of antibody could be detected. In addition, as little as 1 ng of Forssman antigen could be estimated by a binding inhibition experiment.