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Method for screening inhibitors of the toxicity of Bacillus anthracis

Patent ·
OSTI ID:874161
 [1];  [1];  [1]
  1. Los Alamos, NM
+ Show Author Affiliations

The protective antigen (PA) of Bacillus anthracis is integral to the mechanism of anthrax poisoning. The cloning, expression and purification of a 32 kDa B. anthracis PA fragment (PA32) is described. This fragment has also been expressed as a fusion construct to stabilized green fluorescent protein (EGFP-PA32). Both proteins were capable of binding to specific cell surface receptors as determined by fluorescent microscopy and a flow cytometric assay. To confirm binding specificity in the flow cytometric assay, non-fluorescent PA83 or PA32 was used to competitively inhibit fluorescent EGFP-PA32 binding to cell receptors. This assay can be employed as a rapid screen for compounds which disrupts binding of PA to cells. Additionally, the high intracellular expression levels and ease of purification make this recombinant protein an attractive vaccine candidate or therapeutic treatment for anthrax poisoning.

Research Organization:
University of California; Los Alamos National Laboratory (LANL), Los Alamos, NM
Sponsoring Organization:
USDOE
DOE Contract Number:
W-7405-ENG-36
Assignee:
The Regents of the University of California (Los Alamos, NM)
Patent Number(s):
US 6329156
Application Number:
09/273,839
OSTI ID:
874161
Country of Publication:
United States
Language:
English

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