Ultrasensitive Detection of Toxins Using Immunoassay Amplification. Phase 1

摘要

The objective of this Phase I effort, was to develop an amplified immunoassaywhich could detect botulinal neurotoxins A, B, E and F at concentrations equivalent to the mouse toxicity assay (i.e. 5 pg/ml). Reasons for doing this included the likelihood that such an assay would be much shorter than the mouse toxicity (hours rather than days), could be developed for field use, and would be substantially more economical than mouse bioassay. The approach which was to be applied to this problem, was the amplified immunoassay system developed by Elcatech. This system is based on the ultrasensitive measurement of coagulation-activating proteases by generation of a solid-phase enzyme-labeled fibrin matrix (enzyme-linked coagulation assay, or ELCA). When a coagulation-activating protease is attached to an antigen or antibody, then the very sensitive detection of this labeled conjugate permits the very sensitive detection of the immune complexes formed in various assay protocols (ELISA-ELCA). The general principle of this immunoassay is reflected in the Figure below. RA 1; Botulinum toxin; Detection; Assay; Amplification.