A monoclonal antibody-based elisa for the hedgehog inhibitors cyclopamine and cyclopamine-KAAD

In the late 1960's cyclopamine was isolated from the plant Veratrum californicum and identified as the teratogen responsible for craniofacial birth defects including cyclops in the offspring of sheep grazing on mountain ranges in the western United States. More recently, cyclopamine was found to inhibit the hedgehog (Hh) signaling pathway which plays a critical role in embryonic development and is implicated in several types of cancer. Thus, cyclopamine and cyclopamine derivatives have been targeted as potential treatments for certain cancers and other diseases associated with the Hh signaling pathway. A monoclonal antibody-based ELISA was developed to detect and measure cyclopamine and cyclopamine derivatives in biological samples. The limits of detection of the assay for cyclopamine and cyclopamine derivatives were <3.0 pg. This assay was also useful for the detection and measurement of cyclopamine in sera from mice dosed with cyclopamine. The ELISA method described demonstrates the potential of using these techniques for the rapid screening of biological samples for the presence and levels of cyclopamine and cyclopamine derivatives that are Hh inhibitors with anticancer potential.