The United States of America, As Represented by the Secretary, Department of Health and Human Services

Los Alamos National Security, LLC

Anne E Boyer

John R Barr

Conrad Padraig Quinn

Jurgen G Schmidt

Suzanne R Kalb

Hercules Moura

Adrian R Woolfitt

One major problem in diagnosis methods presently available for anthrax is that these methods require several days to produce a result. The only existing treatment for anthrax requires administration soon after infection at a time when patients are exhibiting only mild flu-like symptoms. Thus, a patient may be days beyond the time when treatment would be effective by the time a diagnosis is made. The present invention reduces diagnosis time to as little as four hours providing same day identification of anthrax radically increasing the odds of delivering proper treatment and patient recovery. The rapid identification of anthrax lethal factor activity exhibited by the instant invention is also amenable to in vivo screening protocols for the discovery and development of anthrax vaccines and lethal factor inhibitors. The instant invention isolates and concentrates lethal factor and lethal toxin from nearly any biological sample. By capitalizing on the endopeptidase activity of lethal factor the present invention amplifies output signals producing reliable detection of picomolar concentrations of lethal factor. The instant invention involves novel purification and detection techniques and substrates for rapid, reproducible, and quantitative measurements of anthrax lethal factor in biological samples.

Detection of anthrax pathogenicity factors

The present invention is directed to a method for detecting the presence of clostridial neurotoxins in a sample by mixing a sample with a peptide that can serve as a substrate for proteolytic activity of a clostridial neurotoxin; and measuring for proteolytic activity of a clostridial neurotoxin by a mass spectroscopy technique. In one embodiment, the peptide can have an affinity tag attached at two or more sites.

Mass spectrometry-based methods for detection and differentiation of botulinum neurotoxins

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Atlanta, GA, United States of America

Anne E Boyer