LUMINESCENCE DETECTION REAGENTS FOR GLYCOLIPIDS

ALTERATIONS IN SURFACE CARBOHYDRATES IN GLYCOLIPIDS AND GLYCOPROTEINS OF TUMOR CELLS RELATIVE TO NORMAL CELLS IS A COMMON FEATURE OF MALIGNANT DISEASES. IMMUNOSTAINING PROCEDURES DEVELOPED FOR DETECTING GLYCOLIPID ANTIGENS WITH TUMOR-SPECIFIC ANTIBODIES REPRESENT IMPORTANT NEW TECHNIQUESFOR IMMUNODIAGNOSTICS AND IMMUNOTHERAPEUTICS. THE CURRENT METHODS FOR THESE DIAGNOSTIC TESTS RELY ON REAGENTS THAT ARERADIOACTIVE OR ENZYME LINKED. IN THESE STUDIES, A 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 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. THIS RESEARCH HAS THREE SPECIFIC AIMS. FIRST, 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 DETECT 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 COLORECTAL CARCINOMA-SPECIFIC MONOCLONAL ANTIBODY, CA 19-9. THE ASSAYS AND REAGENTS DEVELOPED WILL HAVE IMPORTANT APPLICATIONS IN BOTH BASIC RESEARCH AND CLINICAL LABORATORIES INVOLVED IN THE ANALYSIS OF COMPLEX CARBOHYDRATES.