Other

Synaptic Research, LLC

US Government As Represented by the Secretary of the Army

Kincaid, Randall L.

Randall L Kincaid

George A Oyler

Jeffrey A Lyon

Paul S Fishman

Yien Che Tsai

Yen Che Tsai

A system and method for identifying aneurotoxin inhibitor employing aneurotoxin substrate complex having a peptide substrate, preferably SNAP-25, a reporter domain on one side of said peptide substrate and an immobilization domain on the opposite side of said peptide substrate. Theneurotoxin inhibitor is identified by its ability to decrease the relative amount of cleaved complex, detected through measuring a decrease in complex bound to a solid support. The method of the present invention also utilizes novel cells that express aneurotoxin substrate complex. Also provided are novel stable cell lines that express thetoxin substrate complex and viral vectors capable of efficiently expressing an active light chain of the BoNT within mammalian cells.

Methods for identifying inhibitors of botulinum neurotoxins

A system and method for identifying a botulinum neurotoxin inhibitor employing a botulinum neurotoxin substrate complex having a peptide substrate, preferably SNAP-25, a reporter domain on one side of said peptide substrate and an immobilization domain on the opposite side of said peptide substrate. The botulinum neurotoxin inhibitor is identified by its ability to decrease the relative amount of cleaved complex, detected through measuring a decrease in complex bound to a solid support. The method of the present invention also utilizes novel cells that express a botulinum neurotoxin substrate complex. The methods of the present invention are adapted for cell based screening to monitor the catalytic activity of a BoNT in living cells and to identify molecules that inhibit the catalytic activity of a BoNT in living cells. Also provided are novel stable cell lines that express the botulinum toxin substrate complex and viral vectors capable of efficiently expressing an active light chain of the BoNT within mammalian cells.

A system and method for identifying aneurotoxin inhibitor employing aneurotoxin substrate complex having a peptide substrate, preferably SNAP-25, a reporter domain on one side of said peptide substrate and an immobilization domain on the opposite side of said peptide substrate. Theneurotoxin inhibitor is identified by its ability to decrease the relative amount of cleaved complex, detected through measuring a decrease in complex bound to a solid support. The method of the present invention also utilizes novel cells that express aneurotoxin substrate complex. The methods of the present invention are adapted for cell based screening to monitor the catalytic activity of a BoNT in living cells and to identify molecules that inhibit the catalytic activity of a BoNT in living cells. Also provided are novel stable cell lines that express thetoxin substrate complex and viral vectors capable of efficiently expressing an active light chain of the BoNT within mammalian cells.

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Rockville, MD, United States of America

Randall L Kincaid