Recent data from several labs suggest that intermediates formed during fibril formation, referred to as protofibrils (PFs), may contribute to neuronal injury in Alzheimer's disease (AD). Currently, there are no antibodies in the marketplace that can distinguish these abnormal oligomeric protofibrils from the normal occurring monomeric ABeta peptide. New England Rare Reagents in collaboration with Brigham and Women's Hospital seeks Phase I funding to assist with the development of antibodies to protofibrillular ABetaPeptide mimetics of ABeta intermediate assemblies can be rationally designed and used as immunogens to generate conformationally restricted antibodies. As proof of concept, we have shown previously that cyclized peptides derived from a region within the bovine pfion protein sequence was able to generate conformation-specific antibodies. Following this lead we propose to: (1) Synthesize cyclized peptides derived from the ABeta sequence where Beta sheet associations may form and conjugate with T-cell epitope peptides. (2) Immunize animal with peptide conjugates and screen serum by ELISA for cyclized and un-cyclized peptide specificity. (3) Isolate clones specific for cyclized peptides and generate monoclonal antibody supernatants. (4) Confirmation of antibody specificity to ABeta PFs by Western blotting and immuno-electron microscopy. The commercialization goals for these reagents are to generate monoclonal antibodies that will recognize the Beta-sheet form of ABeta that could be used for research purposes to identify and understand protofibrils pathophysiology in AD or other abnormal protein-folding diseases. Additionally, if protofibrils form in humans, these novel antibodies may be valuable in identifying these assemblies in blood and/or tissues, helping to diagnose early stages of AD. Moreover, if PFs play an important role in the early pathogenesis of AD these antibodies may be useful for passive immunization, or to identify potential peptide vaccine immunogens. For these reasons we strongly feel these antibodies will be of high commercial value if available to the research community or other commercial enterprises. Generation and initial characterization of these antibodies in this phase will be the foundation for further in vitro and in vivo characterization in Phase II, as well as the scale-up of the antibody production
