Detecting prion disease in a sample comprises contacting the sample with a diagnostically effective amount of an antibody having the ability to bind to PrP CTM in situ

摘要

Detecting prion disease in a sample, is new. Detecting prion disease in a sample comprises contacting the sample with a diagnostically effective amount of an antibody having the ability to bind to PrP C-terminal transmembrane anchored form (CTM) in situ, and determining whether the antibody binds specifically to any material in the sample under conditions, where differential binding of the antibody to PrP isoforms associated with prion disease occurs, where antibody binding is indicative of prion disease. Independent claims are included for the following: (1) a composition comprising a hybridoma cell culture producing monoclonal antibodies which bind specifically to one conformer of PrP selected from the group consisting of Prp CTM and PrP N-terminal transmembrane anchored form (NTM) and have low or no cross reactivity with a second conformer of PrP; (2) a composition comprising a monoclonal antibody which binds to an antigenic determinant of a PrP conformer on the surface of a neuroblastoma cell and has low or no cross reactivity with non-cell surface antigenic determinants of PrP conformers; (3) a hybridoma that expresses monoclonal antibodies having the same antigenic specificity as that of monoclonal antibodies 7VC or 19B10; (4) monoclonal antibodies derived from the hybridoma; (5) a monoclonal antibody fragment derived from a monoclonal antibody having the binding specificity of the monoclonal antibody cited above; (6) a method for promoting cell differentiation by providing a cell with a sufficient amount of the monoclonal antibody, where the cell undergoes differentiation; (7) a method of inhibiting PrP expression in a cell by providing a cell with a sufficient amount of a monoclonal antibody that binds to an antigenic determinant on PrP NTM but not other conformers of mature PrP, where PrP expression in the cell is inhibited; (8) a method for obtaining conformer-specific monoclonal antibodies by contacting conformer-enriched translation products prepared using a cell-free translation system, with a composition comprising one or more monoclonal antibodies; and detecting binding of the monoclonal antibodies to one or more of the conformers by solution imnmnopreciptiation; (9) a method for screening hybridomas to distinguish conformer-specific mabs using solution immunoprecipitation of radiolabelled translation products, synthesized in the presence of microsomal membranes derived from the endoplasmic reticulum, resulting in enrichment for one conformer vs another; (10) a specific antibody (7VC) that recognizes priori protein in a copper-dependent fashion; (11) a specific antibody (19B10) that recognizes SecPrP; and (12) a specific antibody (SS) that recognizes the signal sequence of PrP.