Abstract: Peptide sequences that specifically bind infectious prion protein for the generation of antibodies and therapeutic agents are disclosed herein.

Abstract: Peptide sequences that specifically bind infectious prion protein for the generation of antibodies and therapeutic agents are disclosed herein.

Abstract: The present invention provides antibodies that specifically bind with a high degree of binding affinity to a native ungulate PrPC and/or a denatured ungulate PrPSc, but not to a native ungulate PrPSc. Preferred antibodies find native bovine PrPC and treated PrPSc but not native bovine PrPSc and can be used in an assay to determine if a sample is infected with infectious prions, i.e. PrPSc.

Abstract: An assay method is disclosed which isolates and detects the presence of a disease related conformation of a protein (e.g., PrPSc) present in a sample also containing the non-disease related conformation of the protein (e.g., PrPSc). The sample is treated (e.g., contacted with protease) in a manner which hydrolyzes the disease related conformation and not the non-disease related conformation. The treated sample is contacted with a binding partner (e.g., a labeled antibody which binds PrPSc) and the occurrence of binding provides and indication that PrPSc is present. Alternatively the PrPSc of the treated sample is denatured (e.g., contacted with guanadine) or unfolded. The unfolded PrPSc is contacted with a binding partner and the occurrence of binding indicates the presence of PrPSc in the sample.

Abstract: The invention comprises a formulation and a method which uses the formulation. The formulation is comprised of an aqueous or alcohol solvent having therein (1) a detergent such as SDS; (2) a weak acid such as acetic acid; and (3) a chemical modification reagent such as hydrogen peroxide. The formulation can be modified to substitute other detergents for the SDS, other acids for the acetic acid and other oxidants for the peroxide provided the substitute results in a total formulation which completely inactivates the infectivity of infectious proteins such as prions in a relatively short period of time (e.g. less than two hours) and under relatively mild temperatures (e.g. 60° C. or less).

Abstract: An assay method is disclosed which isolates and detects the presence of a disease related conformation of a protein (e.g., PrPSc) present in a sample also containing the non-disease related conformation of the protein (e.g., PrPC). The sample is treated (e.g., contacted with protease) in a manner which hydrolyzes the disease related conformation and not the non-disease related conformation. The treated sample is contacted with a binding partner (e.g., a labeled antibody which binds PrPSc) and the occurrence of binding provides and indication that PrPSc is present. Alternatively the PrPSc of the treated sample is denatured (e.g., contacted with guanadine) or unfolded. The unfolded PrPSc is contacted with a binding partner and the occurrence of binding indicates the presence of PrPSc in the sample.

Abstract: Infectious proteins such as prions present in a sample are amplified by adding a recombinant form (or portion thereof) of the infectious protein to the sample. The sample with the recombinant protein therein is maintained under cell free conditions which promote amplification for 20 hours or less and then assayed for the infectious protein.

Abstract: A syringe type tissue extraction device is disclosed. The device comprises a sharp, cylindrical metal cuff at one end of a transparent tube which is pushed into brain tissue. Once inserted a plunger is pulled back and by observation through the transparent tube a determined amount of tissue is extracted. The device is then used to place the sample in a single homogenization tube or to punch through a seal above a well of a 96 well plate and the tissue in the tube is inserted into the well. Each well (or the single tube) of the plate holds beads and once tissue is inserted in all the wells the plate is shaken to homogenize the tissue in each well in order to prepare the sample for assaying.

Abstract: The invention provides prion protein standards for use as reference materials for prion detection. The standard may be species specific, i.e. the standard is comprised of a preparation for detection of a single strain prion or it may be prepared to allow detection of multiple prion strains simultaneously. The invention also provides methods of preparing the prion protein standards using a group of non-human host mammals which have their genome manipulated with respect to genetic material related to a PrP gene such that the mammals are susceptible to infection with a prion which generally only infects an animal which is genetically diverse from the host.

Abstract: Antibodies are disclosed which specifically bind to native PrPSc in situ. Preferred antibodies bind only to the native PrPSc of a particular species e.g., human, cow, sheep, pig, etc. Particularly preferred antibodies bind specifically to a particular isoform of human PrPSc. Preferred antibodies of the invention are (1) produced by phage display methodology, (2) bind specifically to native PrPSc, (3) neutralizes the infectivity of prions, (4) bind to PrPSc in situ and (5) bind 50% or more of PrPSc in a liquid flowable sample. Antibodies of the invention can be bound to a substrate and used to assay a sample (which has any PrPSc denatured via proteinase K) for the presence of PrPSc of a specific species which PrPSc is associated with disease. Antibodies which specifically bind to human PrPSc can be labeled and injected carrying out an in vivo diagnostic test to determine if the human is infected with prions associated with disease.

Abstract: Methods are disclosed whereby formulations of molecules are administered rendering cells resistant to infection with infectious proteins such as prions. The formulation preferably comprises a plurality of Fab fragments which (1) recognize and selectively bind to a range of epitopes on the protein of interest, (e.g. epitopes on PrPC) and (2) bind to epitopes which interrupt the chain of events resulting in a change of the protein's conformation to an infectious disease conformation of the protein. The molecules and formulation are also useful in clearing infectious proteins from cells.

Abstract: The present invention provides a novel PrP protein, and nucleic acids encoding this protein, where the PrP protein is characterized in vivo by 1) incomplete glycosylation relative to glycosylation of wild-type PrPC and 2) proper cellular localization, i.e. an ability to be transported to the cell surface. This novel, under-glycosylated PrP, unlike its normal cellular counterpart, can easily be converted into a protease-resistant isoform by incubation with infectious prions. The invention further provides systems for the study of prion disorders and methods of using these systems, e.g. the study of the mechanical processes in progression of prion-mediated disease or the identification of new therapeutic agents for treatment of prion-mediated disorders. In such systems, protease-resistant under-glycosqvated PrP is generated de novo and can be detected by standard immunoblot techniques.

Abstract: The invention is drawn to compositions and methods for inhibiting and treating malformed forms of proteins causing neurodegenerative disease, such as protease resistant prion proteins (PrPSc) and those associated with transmissible spongiform encephalopathies (TSEs). The compounds disclosed herein can be present as racemic mixtures or as compositions consisting essentially only of the optically active isomer in a higher percentage amount e.g. 60% or more, 70% or more, 80% or more, 90% or more or 100% of the optically active isomer and specifically the dextrorotary isomer of quinacrine.