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: 0301 basic medicine; Prions; Science; Blotting, Western; 10208 Institute of Neuropathology; Antibody Affinity; Immunoglobulin Variable Region; 610 Medicine & health; 10071 Functional Genomics Center Zurich; 1100 General Agricultural and Biological Sciences; Cross Reactions; Peptide Mapping; Antibodies; Epitopes; Mice; 03 medical and health sciences; 1300 General Biochemistry, Genetics and Molecular Biology; Animals; Humans; Immunoprecipitation; Enzyme-Linked Immunoassays; Immunohistochemistry Techniques; 1000 Multidisciplinary; Q; R; Antibodies, Monoclonal; Surface Plasmon Resonance; Flow Cytometry; Immunohistochemistry; Recombinant Proteins; 3. Good health; Immunoglobulin Isotypes; 570 Life sciences; biology; Medicine; U7 Systems Biology / Functional Genomics; Epitope Mapping; Cloning; Research Article

PLoS ONE, 3 (12)<br/>PrPSc, a misfolded and aggregated form of the cellular prion protein PrPC, is the only defined constituent of the transmissible agent causing prion diseases. Expression of PrPC in the host organism is necessary for prion replication and for prion neurotoxicity. Understanding prion diseases necessitates detailed structural insights into PrPC and PrPSc. Towards this goal, we have developed a comprehensive collection of monoclonal antibodies denoted POM1 to POM19 and directed against many different epitopes of mouse PrPC. Three epitopes are located within the N-terminal octarepeat region, one is situated within the central unstructured region, and four epitopes are discontinuous within the globular C-proximal domain of PrPC. Some of these antibodies recognize epitopes that are resilient to protease digestion in PrPSc. Other antibodies immunoprecipitate PrPC, but not PrPSc. A third group was found to immunoprecipitate both PrP isoforms. Some of the latter antibodies could be blocked with epitope-mimicking peptides, and incubation with an excess of these peptides allowed for immunochromatography of PrPC and PrPSc. Amino-proximal antibodies were found to react with repetitive PrPC epitopes, thereby vastly increasing their avidity. We have also created functional single-chain miniantibodies from selected POMs, which retained the binding characteristics despite their low molecular mass. The POM collection, thus, represents a unique set of reagents allowing for studies with a variety of techniques, including western blotting, ELISA, immunoprecipitation, conformation-dependent immunoassays, and plasmon surface plasmon resonance-based assays.<br/>ISSN:1932-6203<br/>