Ma Hongzheng, Zhou Bin, Kim YoungSoo, Janda Kim D
Department of Chemistry, The Skaggs Institute for Chemical Biology, Worm Institute for Research and Medicine (WIRM), The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Toxicon. 2006 Jun 15;47(8):901-8. doi: 10.1016/j.toxicon.2006.03.003. Epub 2006 Mar 27.
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 3h 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.
基于一种11聚体环肽(称为C11-019)开发了一种A型肉毒杆菌神经毒素(BoNT/A)ELISA检测系统,该环肽是通过肽噬菌体展示技术鉴定出来的。该检测采用夹心形式,使用附着在聚(马来酸乙烯酯)(PEMA)基质上的C11-019环肽作为捕获阶段,抗BoNT/A多克隆抗体作为检测阶段。报告的结果表明,C11-019肽-聚合物可以特异性结合BoNT/A,在检测缓冲液以及多种体液和食品中对其他检测的血清型无交叉反应。当使用高灵敏度化学发光底物时,在3小时内可轻松实现1 pg/mL的检测,线性范围为0.1-1 ng/mL。这些结果表明,一种基于肽-聚合物的廉价捕获ELISA系统可用于快速、灵敏且高度特异性的BoNT检测。