The POM Monoclonals: A Comprehensive Set of Antibodies to Non-Overlapping Prion Protein Epitopes

• DOI: 10.1371/journal.pone.0003872
• Publication Date: 2008-12-05T22:03:50Z
• Authors: Magdalini Polymen...; Rita Moos; Mike Scott; Christina Sigurdson; Yong-zhong Shi; Bill Yajima; Iva Hafner-Bratkovič; Roman Jerala; Simone Hornemann; Kurt Wuthrich; Anne Bellon; Martin Vey; Graciela Garen; Michael N. G. James; Nat Kav; Adriano Aguzzi
• Keywords: Epitope mapping; Immunoprecipitation; Avidity; Paratope; Conformational epitope

PrPSc, a misfolded and aggregated form of the cellular prion protein PrPC, is only defined constituent transmissible agent causing diseases. Expression PrPC in host organism necessary for replication neurotoxicity. Understanding diseases necessitates detailed structural insights into PrPSc. Towards this goal, we have developed comprehensive collection monoclonal antibodies denoted POM1 to POM19 directed against many different epitopes mouse PrPC. Three are located within N-terminal octarepeat region, one situated central unstructured four discontinuous globular C-proximal domain Some these recognize that resilient protease digestion Other immunoprecipitate but not A third group was found both PrP isoforms. latter could be blocked with epitope-mimicking peptides, incubation an excess peptides allowed immunochromatography Amino-proximal were react repetitive epitopes, thereby vastly increasing their avidity. We also created functional single-chain miniantibodies from selected POMs, which retained binding characteristics despite low molecular mass. The POM collection, thus, represents unique set reagents allowing studies variety techniques, including western blotting, ELISA, immunoprecipitation, conformation-dependent immunoassays, plasmon surface resonance-based assays.