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Applied and Environmental Microbiology, February 2008, p. 818-822, Vol. 74, No. 3
0099-2240/08/$08.00+0     doi:10.1128/AEM.01244-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Development and Implementation of a Single-Chain Fv Antibody for Specific Detection of Bacillus anthracis Spores{triangledown}

A. Mechaly, E. Zahavy, and M. Fisher*

Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 74100, Israel

Received 5 June 2007/ Accepted 7 October 2007

A single-chain Fv (scFv) antibody was developed and applied for efficient and specific detection of Bacillus anthracis spores. The antibody was isolated from a phage display library prepared from spleens of mice immunized with a water-soluble extract of the outer membrane of the B. anthracis spore (exosporium). The library (7 x 106 PFU) was biopanned against live, native B. anthracis ATCC {Delta}14185 spores suspended in solution, resulting in the isolation of a unique soluble scFv antibody. The antibody was affinity purified and its affinity constant (3 x 108 ± 1 x 108 M–1) determined via flow cytometry (FCM). Preliminary characterization of scFv specificity indicated that the scFv antibody does not cross-react with representatives of some phylogenetically related Bacillus spores. The potential use of scFv antibodies in detection platforms was demonstrated by the successful application of the soluble purified scFv antibody in enzyme-linked immunosorbent assays, immunofluorescence assays, and FCM.

* Corresponding author. Mailing address: Department of Infectious Diseases, Israel Institute for Biological Research, P.O. Box 19, Ness-Ziona 74100, Israel. Phone: 972-89381426. Fax: 972-8938174. E-mail: morlyf{at}iibr.gov.il

{triangledown} Published ahead of print on 26 October 2007.

Applied and Environmental Microbiology, February 2008, p. 818-822, Vol. 74, No. 3
0099-2240/08/$08.00+0     doi:10.1128/AEM.01244-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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