Alteration in surface carbohydrates in glycolipids and glcoproteins of tumor cells relative to normal cells is a common feature of malignant diseases. Immunostaining procedures developed specifically for detecting glycolipid antigens with tumor specific antibodies represent important new techniques for immunodiagnostics and immunotherapeutics. The current methods for these diagnostic tests rely on reagents that are radioactive or that are enzyme-linked. In the studies proposed a new and general method will be developed for assaying glycolipids on solid supports that is more rapid and 3 to 6 orders of magnitude more sensitive than existing techniques and that does not utilize radioisotopes. This new technology exploits the remarkable sensitivity of the recombinant, bioluminescent protein, aequorin, and the high affinity interactions of streptavidin and biotin. The work described in this proposal has three specific aims. Fist, assays will be established using luminescence detection reagents (LDR's) for assaying Forssman glycolipid, a human tumor antigen, in microtiter plates. Second, this assay will be adapted to detecting glycolipids separated by thin layer chromatography. Third, an assay will be developed for detecting the sialyl-Le a antigen in complex mixtures using LDR in combination with the colorectalcarcinoma specific monoclonal antibody CA 19-9. The assays and reagents developed during this study will have important applications in both basic research and clinical laboratories involved in analysis of complex carbohydrates.
