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Clinical and Vaccine Immunology, June 2006, p. 671-677, Vol. 13, No. 6
1071-412X/06/$08.00+0     doi:10.1128/CVI.00023-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Detection of Anthrax Toxin in the Serum of Animals Infected with Bacillus anthracis by Using Engineered Immunoassays

Robert Mabry,1 Kathleen Brasky,2 Robert Geiger,2 Ricardo Carrion Jr.,3 Gene B. Hubbard,2 Stephen Leppla,4 Jean L. Patterson,3 George Georgiou,5,6* and B. L. Iverson1,6*

Institute for Cellular and Molecular Biology, University of Texas at Austin, Texas,1 Department of Comparative Medicine, Southwest Foundation for Biomedical Research, San Antonio, Texas,2 Department of Virology and Immunology, Southwest Foundation for Biomedical Research, San Antonio, Texas,3 Bacterial Toxins and Therapeutics Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland,4 Department of Chemical Engineering and Biomedical Engineering, University of Texas at Austin, Austin, Texas,5 Department of Chemistry and Biochemistry, University of Texas at Austin, Austin, Texas6

Received 15 December 2005/ Returned for modification 6 March 2006/ Accepted 17 April 2006

Several strategies that target anthrax toxin are being developed as therapies for infection by Bacillus anthracis. Although the action of the tripartite anthrax toxin has been extensively studied in vitro, relatively little is known about the presence of toxins during an infection in vivo. We developed a series of sensitive sandwich enzyme-linked immunosorbent assays (ELISAs) for detection of both the protective antigen (PA) and lethal factor (LF) components of the anthrax exotoxin in serum. The assays utilize as capture agents an engineered high-affinity antibody to PA, a soluble form of the extracellular domain of the anthrax toxin receptor (ANTXR2/CMG2), or PA itself. Sandwich immunoassays were used to detect and quantify PA and LF in animals infected with the Ames or Vollum strains of anthrax spores. PA and LF were detected before and after signs of toxemia were observed, with increasing levels reported in the late stages of the infection. These results represent the detection of free PA and LF by ELISA in the systemic circulation of two animal models exposed to either of the two fully virulent strains of anthrax. Simple anthrax toxin detection ELISAs could prove useful in the evaluation of potential therapies and possibly as a clinical diagnostic to complement other strategies for the rapid identification of B. anthracis infection.

* Corresponding author. Mailing address for B. L. Iverson: University of Texas at Austin, 1 University Station, Austin, TX 78712. Phone: (512) 471-5053. Fax: (512) 471-7963. E-mail: biverson{at}mail.utexas.edu. Mailing address for G. Georgiou: Deparment of Chemical Engineering and Biomedical Engineering, University of Texas at Austin, 1 University Station, Austin, TX 78712. Phone: (512) 471-6975. Fax: (512) 471-7963. E-mail: gg@che.utexas.edu.

Clinical and Vaccine Immunology, June 2006, p. 671-677, Vol. 13, No. 6
1071-412X/06/$08.00+0     doi:10.1128/CVI.00023-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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