Program Director/Principal Investigator (Last, First, Middle): Vamum, Susan Marie PROJECT SUMMARY (See instructions): There are a number of toxins, includingbotulinumneurotoxin, ricin, shiga toxin and Staphylococcus enterotoxin B (SEB),that have either been used as bioterrorism agents or are considered a bioterrorism threat because of their extreme toxicity or ease of availibility. Furthermore, several of these toxins (botulinum neurotoxin, ricin) work at low concentrations, requiringhighlysensitive assays for detection. Additionally, it is important to distuiguish between active and inactive toxin because of the possibilitythat genetically engineered toxins, consisting of the enzymatic portion of the toxin and a bindingdomain of another protein, can be used as the bioweapon agent. Consequently,our long-term goal is to develop diagnostic assays for toxin detection and differentiation that address these dual requirements of high sensitivity and determination of toxin activity. Our objective in this proposal is to develop a platform for toxin detection based upon a combination of two analytical approaches: 1) molecular recognition based upon ELISA microarrays and 2) enzymaticactivity assays for toxin activity. We propose that this combined platform will provide a highly sensitive and specfic assay for the detection of these toxins.
The specific aims are: 1). Developan ELISA microarray for the sensitive and quantitative detection of botulinum neurotoxins (serotypes A,B,C,D,E,and F), ricin toxin, shiga toxin and Staphylococcal enterotoxin B. 2). Functionalactivity assays specific for botulinumneurotoxins, ricin, and shiga toxin will be developed. 3). A single platform, comprised of the ELISA microarray and the activity assay will be developed, allowing for the sensitive and specificdetection and the determination of functional activity of,botulinumneurotoxins,ricin, Shiga toxin and Staphylococcal enterotoxin B in clinical samples. The combined assay platformwill be used to analyze clinical samples, includingsera, nasal swabs and stool samples, and for the determination of the sensitivity and specificityof the assays.

Public Health Relevance

In the event of a bioweapon attack, it will be critical to rapidly determine which bioweapon agent is responsible for the attack in order to give the appropiate medical care. Our objective in this proposal is to develop a sensitive and specific assay for the detection of the following toxins: botulinum neurotoxin, ricin toxin, shiga toxin and Staphylococcus enterotoxin B. PROJECT/

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01AI081895-05
Application #
8495216
Study Section
Special Emphasis Panel (ZAI1-MMT-M (J1))
Program Officer
Hall, Robert H
Project Start
2009-07-15
Project End
2014-06-30
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
5
Fiscal Year
2013
Total Cost
$449,551
Indirect Cost
$210,695
Name
Battelle Pacific Northwest Laboratories
Department
Type
DUNS #
032987476
City
Richland
State
WA
Country
United States
Zip Code
99352
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Zhang, Yanfeng; Gardberg, Anna S; Edwards, Thomas E et al. (2013) Structural insights into the functional role of the Hcn sub-domain of the receptor-binding domain of the botulinum neurotoxin mosaic serotype C/D. Biochimie 95:1379-85
Zhang, Yanfeng; Varnum, Susan M (2012) The receptor binding domain of botulinum neurotoxin serotype C binds phosphoinositides. Biochimie 94:920-3
Zhang, Yanfeng; Buchko, Garry W; Qin, Ling et al. (2011) Crystal structure of the receptor binding domain of the botulinum C-D mosaic neurotoxin reveals potential roles of lysines 1118 and 1136 in membrane interactions. Biochem Biophys Res Commun 404:407-12
Zhang, Yanfeng; Buchko, Garry W; Qin, Ling et al. (2010) Structural analysis of the receptor binding domain of botulinum neurotoxin serotype D. Biochem Biophys Res Commun 401:498-503