A Chemically Synthesized Capture Agent Enables the Selective, Sensitive, and Robust Electrochemical Detection of Anthrax Protective Antigen
Abstract
We report on a robust and sensitive approach for detecting protective antigen (PA) exotoxin from Bacillus anthracis in complex media. A peptide-based capture agent against PA was developed by improving a bacteria display-developed peptide into a highly selective biligand through in situ click screening against a large, chemically synthesized peptide library. This biligand was coupled with an electrochemical enzyme-linked immunosorbent assay utilizing nanostructured gold electrodes. The resultant assay yielded a limit of detection of PA of 170 pg/mL. (2.1 pM) in buffer, with minimal sensitivity reduction in 1% serum. The powdered capture agent could be stably stored for several days at 65 °C, and the full electrochemical biosensor showed no loss of performance after extended storage at 40 °C. The engineered stability and specificity of this assay should be extendable to other cases in which biomolecular detection in demanding environments is required.
Additional Information
© 2013 American Chemical Society. Publication Date (Web): September 24, 2013. Published In Issue October 22, 2013. Article ASAP October 08, 2013. Just Accepted Manuscript September 24, 2013. Received: August 16, 2013. Accepted: September 24, 2013. This research was funded primarily provided by the Institute for Collaborative Biotechnologies through grant W911NF-09-0001 from the U.S. Army Research Office. Research is supported in part by appointments (M.B.C.) to the U.S. Army Research Laboratory Postdoctoral Fellowship Program administered by the Oak Ridge Associated Universities through a contract with the U.S. Army Research Laboratory. The content of the information does not necessarily reflect the position or the policy of the Government, and no official endorsement should be inferred. The research was partially supported by the National Research Foundation of Korea Grant funded by the Korean Government (MEST) (2011-0028861), and a grant from the Institute of Medical System Engineering (iMSE) of GIST, Republic of Korea. The authors would like to thank J. M. Kogot, D. A. Sarkes, and P. M. Pellegrino for their contributions towards the previous discovery of anti-protective antigen peptide sequences. The following reagents were obtained through the NIH Biodefense and Emerging Infections Research Resources Repository, NIAID, NIH: Anthrax Protective Antigen (PA), recombinant from E. coli, NR-3780.Errata
There are no corrections to the actual article file; we are only modifying Supplementary Figure S10, part C. The original Figure S10C is incorrect and was a simple mistake. The new Figure S10C now represents a more complete analysis. None of the conclusions of the main text are changed.Attached Files
Supplemental Material - nn404296k_si_001.pdf
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Additional details
- Eprint ID
- 42991
- Resolver ID
- CaltechAUTHORS:20131212-131040336
- W911NF-09-0001
- Army Research Office (ARO)
- MEST-2011-0028861
- National Research Foundation of Korea
- Institute of Medical System Engineering
- Created
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2013-12-12Created from EPrint's datestamp field
- Updated
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2021-11-10Created from EPrint's last_modified field