ELCATECH HAS DEVELOPED A NEW TECHNIQUE FOR THE IMMUNOASSAY OF CLOTTING FACTORS.

ELCATECH HAS DEVELOPED A NEW TECHNIQUE FOR THE IMMUNOASSAY OF CLOTTING FACTORS. THE TECHNIQUE IS SIMILAR TO THE ELISA METHODOLOGY IN THAT ENZYME-LABELED DERIVATIVES OF ANTIGENS AND ANTIBODIES ARE USED TO DETECT IMMUNE COMPLEXES. IT DIFFERS FROM MOST ELISA METHODS IN THAT IT USES AN ACTIVATOROF COAGULATION (RUSSELL'S VIPER VENOM FACTOR X ACTIVATOR OR RVV-XA) AS THE LABELING ENZYME AND A UNIQUE CLOTTING ASSAY, ENTITLED ENZYME-LINKED COAGULATION ASSAY (ELCA), AS THE DETECTION SYSTEM. ELCA USES PEROXIDASE-LABELED FIBRINOGEN IN SOLUTION AND UNLABELED FIBRINOGEN TOGETHER AS A SUBSTRATEFOR THROMBIN, WHICH IS GENERATED IN THE CLOTTING CASCADE. THROMBIN CLEAVES BOTH FORMS TO FIBRIN AND CONSEQUENTLY PEROXIDASE-FIBRIN IS BOUND TO THE SOLID PHASE. USED AS A DETECTION SYSTEM FOR RVV-XA, THIS ASSAY CAN MEASURE AS LITTLE AS 50 FEMTOGRAMS/ML OF RVV-XA, WHICH IS MORE THAN THREE ORDERS OF MAGNITUDE LOWER THAN THE DETECTION LIMIT FORPEROXIDASE. ANTIGENS AND ANTIBODIES CAN BE LABELED WITH THIS ENZYME; THESE CONJUGATES CAN THEN BE USED FOR COMPET- ITIVE IMMUNOASSAYS WITH HIGH SENSITIVITY. BY COMBINATION OFTHE SPECIFICITY OF MONOCLONAL ANTIBODIES FOR FACTORS OF INTEREST (II,VII,IX,X,AND PROTEIN C) WITH THIS SENSITIVE DETECTION SYSTEM, RESEARCHERS WILL ASSAY THREE IMPORTANT PARAMETERS OF INTEREST IN THROMBOSIS AND HEMOSTASIS, THAT IS, ACTIVATED FACTOR X, CLOTTING FACTOR ANTIGEN LEVELS, AND COMPLEXES OF FACTOR XA WITH ANTITHROMBIN III. THESE IMMUNO-ASSAYS, IN CONJUNCTION WITH THE SENSITIVE FUNCTIONAL DETECT-ION OF CLOTTING FACTORS BY ELCA, WILL PROVIDE A COMPLETE SYSTEM FOR THE DETECTION OF COMPONENTS OF INTEREST IN HEMOSTASIS AND THROMBOSIS.