ULTRASENSITIVE DETECTION OF TOXINS USING IMMUNOASSAY AMPLIFICATION

THE INABILITY TO DETECT TOXINS WITH ADEQUATE SENSITIVITY IS ONE OF THE PRINCIPAL REASONS THAT THEY ARE POTENTIALLY THREATENING, EITHER DUE TO "NATURAL" EXPOSURE OR DELIBERATE AND HOSTILE USE OF THESE AGENTS BY AN ADVERSARY. THIS PROPOSAL OUTLINES A WAY THAT THE DETECTION OF THESE TOXINS CAN BE MADE MUCH MORE SENSITIVE WITHOUT USE OF EXPENSIVE EQUIPMENT OR HIGHLY TRAINED PERSONNEL FOR THE FINAL PRODUCT. THE BASIS OF THE METHOD LIES IN THE USE OF COAGULATION-ACTIVATING SNAKE VENOM ENZYME CONJUGATES OF ANTIBODIES AND ANTIGENS AND THEIR DETECTION BY A NEW, ULTRASENSITIVE COAGULATION ACTIVATOR ASSAY CALLED ENZYME-LINKED COAGULATION ASSAY, OR ELCA. WE EXPECT THAT WE COULD MEASURE TOXINS WITH 100-1000X HIGHER SENSITIVITY THAN OTHER CURRENT METHODOLOGIES, BASED ON OUR CURRENT SUCCESS IN APPLYING THIS ASSAY TO OTHER ANTIGENS. DURING THIS PHASE I EFFORT, WE WOULD DEMONSTRATE THE POTENTIAL OF THE TECHNOLOGY FOR TOXINS OF INTEREST TO THE ARMY, BY LABELING THESE TOXINS AND/OR ANTIBODIES TO THEM, AND DEMONSTRATING THE SENSITIVITY OF DETECTION USING THE ELCA ASSAY. A VARIETY OF PROTOCOLS FOR THE ASSAY WOULD BE TESTED, ALL BEING DEPENDENT ON THE ELCA ASSAY FOR FINAL DETECTION.