P2-372: Development of oligomer-specific alpha-synuclein antibodies

Abstract

Aggregates of alpha-synuclein (a-syn) are present as Lewy bodies and Lewy neurites in the brains of patients suffering from dementia with Lewy bodies (DLB) and Parkinson′s disease (PD). Accumulating evidence suggests that oligomers, intermediately sized soluble species of the a-syn aggregation process, are primarily responsible for the neurotoxicity in these disorders. Our aim is to develop oligomer specific monoclonal antibodies as tools for studies on cell cultures and transgenic mice with a-syn pathology. Such antibodies might be useful for diagnostic purposes and for immunotherapy of DLB and PD. Mice were repeatedly immunized subcutaneously with preparations of oligomeric recombinant a-syn. Next, spleen cells were fused with mouse myeloma cells to generate antibody producing hybridomas. Finally, the generated clones were screened, using ELISA, for a-syn antibodies specifically binding to the oligomeric state. A total of five mice (Balb/c) were immunized with oligomeric a-syn preparations. All mice responded to the immunization and antibodies against monomeric, oligomeric and fibrillar a-syn forms were detected in plasma. After generation of hybridomas, some of the clones showed specificity to a-syn oligomers whereas others were more selective for a-syn monomers and fibrils. Ongoing subcloning aims at further isolating IgG-positive clones with a high degree of a-syn oligomer specificity. Preliminary results indicate that we have generated antibodies specific to a-syn oligomers. Further screening and characterization of these antibodies are ongoing.