Chemistry: natural resins or derivatives; peptides or proteins; – Peptides of 3 to 100 amino acid residues
Reexamination Certificate
2005-03-31
2009-12-15
Kam, Chih-Min (Department: 1656)
Chemistry: natural resins or derivatives; peptides or proteins;
Peptides of 3 to 100 amino acid residues
C530S324000, C530S333000, C530S350000, C424S239100, C435S004000, C435S007710
Reexamination Certificate
active
07632917
ABSTRACT:
A system and method for identifying abotulinumneurotoxin inhibitor employing abotulinumneurotoxin 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. Thebotulinumneurotoxin 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 abotulinumneurotoxin 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 thebotulinumtoxin substrate complex and viral vectors capable of efficiently expressing an active light chain of the BoNT within mammalian cells.
REFERENCES:
patent: 5965699 (1999-10-01), Schmidt et al.
patent: 6203794 (2001-03-01), Dolly et al.
patent: 6504006 (2003-01-01), Shine et al.
patent: 6762280 (2004-07-01), Schmidt et al.
patent: 2001/0012833 (2001-08-01), Aoki et al.
patent: 2002/0127247 (2002-09-01), Steward et al.
patent: 2003/0077685 (2003-04-01), Schmidt et al.
patent: 2003/0143650 (2003-07-01), Steward et al.
patent: 2003/0143651 (2003-07-01), Steward et al.
patent: 2004/0072270 (2004-04-01), Fernandez-Salas et al.
patent: 2004/0115727 (2004-06-01), Steward et al.
patent: 2004/0146963 (2004-07-01), Schmidt et al.
patent: 2005/0100973 (2005-05-01), Steward et al.
patent: 2005/0136394 (2005-06-01), Fang et al.
patent: 2005/0153945 (2005-07-01), Bavari et al.
Rossetto et al. Nature vol. 372, 415-416 (Dec. 1994).
Washbourne et al. , FEBS Lett. 418, 1-5 (1997).
Hallis B., et al., Development of Novel Assays forbotulinumType A and B Neurotoxins Based on Their Endopeptidase Activities, Journal of Clinical Microbiology, 34(8):1934-1938, (Aug. 1996).
Wictome M., et al., Dev Biol Stand, Novel assays for the detection ofbotulinumtoxins in foods, 101:141-5 (1999).
Szilagyi M., et al., Toxicon, Development of sensitive colorimetric capture elisas forClostridium botulinumneurotoxin serotypes A and B, 38(3):381-389 (Mar. 2000).
Shone CC., et al., Proteolytic cleavage of synthetic fragments of vesicle-associated membrane protein, isoform-2 bybotulinumtype B neurotoxin, Eur J Biochem, 217(3):965-71 (Nov. 1, 1993).
Ekong Ta, et al., Recombinant SNAP-25 is an effective substrate forClostridium botulinumtype A toxin endopeptidase activity in vitro, Microbiology, 143 ( Pt 10):3337-47 (Oct. 1997).
Wictome M., et al., Development of an In Vitro Bioassay forClostridium botulinumType B Neurotoxin in Foods That Is More Sensitive than the Mouse Bioassay, Applied and Environmental Microbiology, 65(9):3787-3792 (Sep. 1999).
Sharma SK, et al., Methods for detection ofClostridium botulinumtoxin in foods, J Food Prot., (6)1256-63 (Jun. 2005).
Fasshauer, Dirk et al., Conserved Structural Features of the Synaptic Fusion Complex: SNARE Proteins Reclassified as Q- and R- SNAREs, Proc. Natl. Acad. Sci. USA, 1998, 15781-15786, vol. 95.
Steegmaier, Martin et al., Three Novel Proteins of the Syntaxin/SNAP-25 Family, J. Biol. Chem., 1998, 34171-34179, vol. 273, No. 51, The American Society for Biochemistry and Molecular Biology, Inc.
Vaidyanathan, Vadakkanchery V. et al., Proteolysis of SNAP-25 Isoforms bybotulinumNeurotoxin Types A, C, and E: Domains and Amino Acid Residues Controlling the Formation of Enzyme-Substrate Complexes and Cleavage, J. Neurochem., 1999, 327-337, vol. 72, No. 1, Lippincott Williams & Wilkins, Philadelphia.
Hearn, Milton T. W., et al., Applications of Novel Affinity Cassette Methods: Use of Peptide Fusion Handles for the Purification of Recombinant Proteins, J. Mol. Recognit., 2001, 323-369, vol. 14.
Fishman Paul S.
Kincaid Randall L.
Oyler George
Tsai Yen Che
Kam Chih-Min
Morales, Esq. Joseph L.
Whiteford Taylor & Preston LLP
LandOfFree
Methods for identifying inhibitors of botulinum neurotoxins does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Methods for identifying inhibitors of botulinum neurotoxins, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Methods for identifying inhibitors of botulinum neurotoxins will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-4066587