Radioimmunoassay of glycosphingolipids: application for the detection of forssman glycolipid in tissue extracts and cell membranes.
Development of a radioimmunoassay for detecting glycosphingolipids has been difficult, primarily because of transfer of radiolabeled glycolipid antigen to the unlabeled antigen pool. This difficulty has been overcome by the use of a radiolabeled glycolipid-polymer. Thus, an assay system has been developed for measuring picomolar quantities of Forssman hapten glycolipid. This assay is based on competition for rabbit anti-Forssman antibodies between Forssman glycolipid and a radiolabeled Forssman polyacrylic hydrazide polymer. Antigen-antibody complexes are removed quickly and efficiently by binding to formalin-fixed Staphylococcus aureus and subsequent centrifugation. One nanogram of Forssman glycolipid can be readily detected both in plasma membrane preparations and in purified glycolipid fractions. The isoantigenic expression of Forssman glycolipid in human gastrointestinal tissues has been reported previously (Hakomori et al., 1977). Using the radioimmunoassay, the Forssman status of several additional cases has been determined quantitatively.
D000818:Animals; D002462:Cell Membrane / Q000032:analysis; D005577:Forssman Antigen / Q000032:analysis*; D006028:Glycosphingolipids / Q000032:analysis*; D006041:Goats; D006801:Humans; D011863:Radioimmunoassay / Q000379:methods*; D014020:Tissue Extracts / Q000032:analysis*
D006028:Glycosphingolipids; D014020:Tissue Extracts; D005577:Forssman Antigen