Abstract: A method is provided for determining the presence of soluble, misfolded ?-synuclein protein in a biological sample. The method comprises contacting the biological sample with a pre-incubation mixture, the pre-incubation mixture comprising: a monomeric ?-synuclein protein; a buffer composition; a salt; and an indicator, to form an incubation mixture. An incubation cycle is conducted on the incubation mixture in the presence of either a silicon nitride bead or a borosilicate glass bead having a diameter of from about 1 mm to about 5 mm. The method further comprises determining if a detectable amount of misfolded ?-synuclein aggregate is present in the biological sample.

Abstract: Methods and kits are provided for reproducible detection of misfolded aS aggregates in biological fluids and tissue that are less-invasively or non-invasively obtained compared to cerebrospinal fluid, such as skin, olfactory mucosa, saliva, and blood or blood parts.

Abstract: Inert matrices for use with a-synuclein seed amplification assays (“?S-SAA”s) are provided. The inert matrices accurately reflect the absence of misfolded ?S protein when used as a negative control, in the form of no, perceptively low, or delayed ?S substrate self-aggregation, yet will readily permit aggregation of the ?S substrate with seeds when used as a positive control. The inert matrices may be used to screen for ?S-SAA reagent competence. The inert matrices may be used to dilute samples taken from peripheral biological matrices. Finally, the inert matrices may be used as a diluent for serial dilutions of ?S-SAA samples, to enable semi-quantitative versions of ?S-SAAs.

Abstract: An expression vector is provided for production of human alpha-synuclein (?S) protein or a conservative variant thereof that exhibits a decreased tendency to self-aggregate in an ?S seed amplification assay (SAA). The expression vector comprises a nucleic acid sequence coding for human ?S protein or a conservative variant, the nucleic acid sequence comprising codons that have been optimized to produce human ?S protein or a conservative variant when expressed by a host cell such as E. Coli. The codons have been optimized to avoid amino acid misincorporation in the expressed protein. Methods for purification of the expressed protein are also provided.

Abstract: Methods and kits are provided for amplifying and detecting misfolded tau protein from samples, for example, from patients having tauopathies such as Alzheimer's Disease, Progressive Supranuclear Palsy, and the like.

Abstract: An expression vector is provided for production of human alpha-synuclein (?S) protein or a conservative variant thereof that exhibits a decreased tendency to self-aggregate in an ?S seed amplification assay (SAA). The expression vector comprises a nucleic acid sequence coding for human ?S protein or a conservative variant, the nucleic acid sequence comprising codons that have been optimized to produce human ?S protein or a conservative variant when expressed by a host cell such as E. coli. The codons have been optimized to avoid amino acid misincorporation in the expressed protein. Methods for purification of the expressed protein are also provided.

Abstract: Methods and kits are provided for amplifying and detecting misfolded tau protein from samples, for example, from patients having tauopathies such as Alzheimer's Disease, Progressive Supranuclear Palsy, and the like.

Abstract: A method is provided for determining the presence of soluble, misfolded ?-synuclein protein in a biological sample. The method comprises contacting the biological sample with a pre-incubation mixture, the pre-incubation mixture comprising: a monomeric ?-synuclein protein; a buffer composition; a salt; and an indicator, to form an incubation mixture. An incubation cycle is conducted on the incubation mixture in the presence of either a silicon nitride bead or a borosilicate glass bead having a diameter of from about 1 mm to about 5 mm. The method further comprises determining if a detectable amount of misfolded ?-synuclein aggregate is present in the biological sample.

Abstract: An expression vector is provided for production of human alpha-synuclein (?S) protein or a conservative variant thereof that exhibits a decreased tendency to self-aggregate in an ?S seed amplification assay (SAA). The expression vector comprises a nucleic acid sequence coding for human ?S protein or a conservative variant, the nucleic acid sequence comprising codons that have been optimized to produce human ?S protein or a conservative variant when expressed by a host cell such as E. coli. The codons have been optimized to avoid amino acid misincorporation in the expressed protein. Methods for purification of the expressed protein are also provided.

Abstract: A method is provided for determining the presence of soluble, misfolded ?-synuclein protein in a biological sample. The method comprises contacting the biological sample with a pre-incubation mixture, the pre-incubation mixture comprising: a monomeric ?-synuclein protein; a buffer composition; a salt; and an indicator, to form an incubation mixture. An incubation cycle is conducted on the incubation mixture in the presence of either a silicon nitride bead or a borosilicate glass bead having a diameter of from about 1 mm to about 5 mm. The method further comprises determining if a detectable amount of misfolded ?-synuclein aggregate is present in the biological sample.

Abstract: A method is provided for distinguishing between and/or diagnosing Parkinson's disease (PD) or multiple system atrophy (MSA) in a subject who is exhibiting symptoms associated with both PD and MSA. The method comprises: (A) contacting a biological sample obtained from the subject and comprising soluble, misfolded alpha-synuclein (?S) protein with a pre-incubation mixture comprising a monomeric ?S substrate and an indicator to form an incubation mixture; (B) conducting an incubation cycle two or more times on the incubation mixture to form misfolded ?S aggregates; (C) subjecting the incubation mixture to excitation and detecting via indicator fluorescence emission the misfolded ?S aggregates; and (D) diagnosing the subject has having PD or MSA depending on the fluorescence emission intensity. In some aspects, the incubation cycles are conducted in the presence of a bead.

Abstract: Methods and kits are provided for amplifying and detecting ?S proteins from samples, for example, from patients having Parkinson's Disease. For example, a method for determining a presence of a soluble, misfolded ?S protein may include: contacting the sample with a monomeric, folded ?S protein to form an incubation mixture; conducting an incubation cycle two or more times on the incubation mixture effective to form an amplified portion of misfolded ?S protein; incubating the incubation mixture effective to cause misfolding and/or aggregation of at least a portion of the monomeric, folded ?S protein in the presence of the soluble, misfolded ?S protein; physically disrupting the incubation mixture effective to at least partly de-aggregate at least a portion of a misfolded ?S aggregate present; and determining the presence of the soluble, misfolded ?S protein in the sample by detecting at least a portion of the soluble, misfolded ?S protein.

Abstract: A method is provided for distinguishing between and/or diagnosing Parkinson's disease (PD) or multiple system atrophy (MSA) in a subject who is exhibiting symptoms associated with both PD and MSA. The method comprises: (A) contacting a biological sample obtained from the subject and comprising soluble, misfolded alpha-synuclein (?S) protein with a pre-incubation mixture comprising a monomeric ?S substrate and an indicator to form an incubation mixture; (B) conducting an incubation cycle two or more times on the incubation mixture to form misfolded ?S aggregates; (C) subjecting the incubation mixture to excitation and detecting via indicator fluorescence emission the misfolded ?S aggregates; and (D) diagnosing the subject has having PD or MSA depending on the fluorescence emission intensity. In some aspects, the incubation cycles are conducted in the presence of a bead.

Abstract: Methods and kits are provided for amplifying and detecting ?S proteins from samples, for example, from patients having Parkinson's Disease. For example, a method for determining a presence of a soluble, misfolded ?S protein may include: contacting the sample with a monomeric, folded ?S protein to form an incubation mixture; conducting an incubation cycle two or more times on the incubation mixture effective to form an amplified portion of misfolded ?S protein; incubating the incubation mixture effective to cause misfolding and/or aggregation of at least a portion of the monomeric, folded ?S protein in the presence of the soluble, misfolded ?S protein; physically disrupting the incubation mixture effective to at least partly de-aggregate at least a portion of a misfolded ?S aggregate present; and determining the presence of the soluble, misfolded ?S protein in the sample by detecting at least a portion of the soluble, misfolded ?S protein.

Abstract: Methods and kits are provided for amplifying and detecting misfolded proteins from samples, for example, from patients having Alzheimer's Disease, Parkinson's Disease, and the like. For example, a method for determining a presence of soluble, misfolded protein in a sample may include contacting the sample with a monomeric, folded protein to form an incubation mixture; conducting an incubation cycle two or more times effective to form an amplified portion of misfolded protein; incubating the incubation mixture effective to cause misfolding and/or aggregation of at least a portion of the monomeric, folded protein; physically disrupting the incubation mixture effective to break up at least a portion of any protein aggregate present; and determining the presence of the soluble, misfolded protein in the sample by detecting at least a portion of the soluble, misfolded protein. The monomeric, folded protein and the soluble, misfolded protein may exclude prion protein (PrP) and isoforms thereof.

Abstract: Methods and kits are provided for amplifying and detecting misfolded tau protein from samples, for example, from patients having tauopathies such as Alzheimer's Disease, Progressive Supranuclear Palsy, and the like.

Abstract: Methods and kits are provided for amplifying and detecting misfolded proteins from samples, for example, from patients having Alzheimer's Disease, Parkinson's Disease, and the like. For example, a method for determining a presence of soluble, misfolded protein in a sample may include contacting the sample with a monomeric, folded protein to form an incubation mixture; conducting an incubation cycle two or more times effective to form an amplified portion of misfolded protein; incubating the incubation mixture effective to cause misfolding and/or aggregation of at least a portion of the monomeric, folded protein; physically disrupting the incubation mixture effective to break up at least a portion of any protein aggregate present; and determining the presence of the soluble, misfolded protein in the sample by detecting at least a portion of the soluble, misfolded protein. The monomeric, folded protein and the soluble, misfolded protein may exclude prion protein (PrP) and isoforms thereof.

Abstract: Methods and kits are provided for amplifying and detecting A? proteins from samples, for example, from patients having Alzheimer's Disease. For example, a method for determining a presence of a soluble, misfolded A? protein may include contacting the sample with a monomeric, folded A? protein to form an incubation mixture; conducting an incubation cycle two or more times on the incubation mixture effective to form an amplified portion of misfolded A? protein; incubating the incubation mixture effective to cause misfolding and/or aggregation of at least a portion of the monomeric, folded A? protein; physically disrupting the incubation mixture effective to at least partly de-aggregate at least a portion of a misfolded A? aggregate present; and determining the presence of the soluble, misfolded A? protein in the sample by detecting at least a portion of the amplified portion of misfolded A? protein.