A Botulinum neurotoxin serotype A (BoNT/A) ELISA detection system was developed based upon an 11-mer cyclic peptide, termed C11-019, that was identified through peptide phage display technology. The assay employs a sandwich format using the C11-019 cyclic peptide attached to a PEMA (poly(ethylene maleic anhydride)) matrix as the capture phase and anti-BoNT/A polyclonal antibodies as the detection phase. Results reported demonstrate that the C11-019 peptide-polymer can specifically bind to BoNT/A with no cross-reactivity to other serotypes examined in assay buffers and a variety of body fluids and foodstuffs. When a highly sensitive chemiluminescent substrate was engaged, the detection of 1 pg/mL could be readily achieved within 3 h with a linear range of 0.1-1 ng/mL. These results demonstrate that an inexpensive peptide-polymer-based capture ELISA system can be used for rapid, sensitive and highly specific BoNT detection.
Bibliographical noteFunding Information:
This work was supported by the National Institutes of Health (U01 AI061194 and contract number HHSN266200400026C to K.D.J.) and The Skaggs Institute for Chemical Biology. We also thank Dr. Mark S. Hixon for helpful discussions of the data.
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