Development and Application of Single Chain Antibodies for PD Therapy

摘要

In PD the insult to the dopamine (DA) neuron is posited to involve oxidative injury mediated by mitochondrial respiratory abnormalities and through participation of oxidative adducts made onto DA and presynaptic target proteins such as alpha-synuclein. The misfolding of alpha-synuclein engendered by oxidative adduct formation is hypothesized to be a critical participating process in Lewy Body formation and dopamine neuron compromise and death. Our central hypothesis purports that protein aggregates forming within dopaminergic neurons are seeded and require misfolded alpha-synuclein and that these aggregates are cytotoxic thereby contributing directly to neuron death. Thus targeting alpha-synuclein protein misfolding will enable the development of effective therapy. The main goal of this application is to identify and characterize humanized single chain antibodies (scFvs) that recognize structural epitopes on alpha-synuclein and utilize these scFvs to affenuate the pathology associated with alpha-synuclein misfolding. Thusfar we have expressed, purified and generated altered conformers of alpha-synuclein; screened for, identified and purified synuclein conformers specific scFvs. We have begun the characterization of these synuclein-specific scFvs in tissue culture and animal models of alpha-synuclein overexpression.