, 2007). BoNT is produced as a dichain polypeptide that is then cleaved into a ~ 100 kDa heavy

chain (HC) and a ~ 50 kDa light chain (LC) (Montal, 2010). While the HC facilitates entry of the toxin into neurons by endocytosis, the LC is a metallopeptidase that cleaves soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), inhibiting acetylcholine release and resulting in flaccid muscle paralysis ( Montal, 2010 and Schiavo et al., 2000). In humans, a lethal dose intravenously or intramuscularly is estimated at 1–2 ng/kg body weight; orally at 1 μg/kg and 10–12 ng/kg by inhalation ( Arnon et al., 2001). Given their potency, BoNTs have been employed as INCB018424 therapeutics, as tools in basic science research, and as weapons of biological warfare (Arnon et al., 2001 and Shukla and Sharma, 2005). The gold standard of

detection of BoNTs is the mouse bioassay, which can detect as little as 10 pg/mL of toxin (Sharma and Whiting, 2005). However, the assay requires several days to complete, large numbers of animals and can only be performed at a select number of laboratories in the United States. To determine the serotype, a second, independent neutralization assay is required. Ribociclib In the event of a suspected BoNT contamination event, the mouse bioassay, while extremely sensitive, does not meet the needs of emergency responders. Therefore a rapid, sensitive, and selective BoNT diagnostic test that can be field deployed could be used to address suspect BoNT contamination. A number of in vitro assays to detect BoNTs, including ELISA kits, PCR-based methods and assays based on the enzymatic activity of the toxin’s light chain have been developed ( Chao et al., 2004, Wictome et al., 1999 and Shone et al., 1985). While some of these methods have comparable sensitivity to the mouse bioassay, they still require trained personnel and specialized equipment. In contrast, lateral flow devices (LFDs) are simple, low cost

alternatives Buspirone HCl that can be easily deployed in the field and do not require specialized training to operate or to interpret the results. LFDs can be read without optical detection systems, are compact, and on average have a long shelf life ( Posthuma-Trumpie et al., 2009, Warsinke, 2009 and Ngom et al., 2010). While these devices typically have less sensitivity than ELISA formats, they do offer a method for rapid, simple assessment of potential BoNT contamination to a multitude of personnel. Our laboratory has developed several high affinity monoclonal antibodies (mAbs) that selectively recognize the BoNT/A and /B serotypes. MAb F1-2, which recognizes the N-terminus of the heavy chain of BoNT/A, has been extensively characterized and effectively employed as a capture antibody in a sandwich ELISA (Scotcher et al., 2009 and Stanker et al., 2008). We have also previously described MCS-6-27, a BoNT/B-specific mAb that binds the carboxyl portion of the HC and can be used as a capture antibody in a sandwich ELISA (Scotcher et al.