Evolving a Human Enzyme into a Catalytic Bioscavenger

Recent changes in the security situation facing citizens in the United States and military personnel abroad have greatly increased the threat that chemical weapons will be used against American forces and American civilians in the field as well as at home. Many third world countries and terrorist groups have the capability of producing and delivering chemical agents thus posing a substantial and serious threat to the armed forces of the United States. There is a great need for protection of military personnel from unexpected nerve agent exposure during warfare, as well as protection of rescuers and medical personnel from exposure after assessing that a nerve agent attack has occurred. The objectives of this Phase I proposal are to develop an expression system to express a highly diversified mutation library and to validate a high-throughput assay for the molecular evolution of hBchE. The success of Phase I will establish tools for molecular evolution efforts in Phase II involving multiple rounds of large-scale functional screening.