Methods and compositions for tauopathy diagnosis and treatment
AssigneeChildren's Medical Center Corporation
InventorsJudith AJ Steen, Hanno Steen, Waltraud Mair, Jan Muntel, Shaojun Tang
Life of this patent
Estimating the term…
Abstract
This disclosure relates to methods of determining the amount of post translational modification (PTM) associated with one or more tau peptide fragments of a tau protein in a sample, and methods of evaluating a subject for having a tauopathy, the methods comprising, in part, determining the amount of post translational modification (PTM) associated with one or more tau peptide fragments of a tau protein in a sample, and comparing the amount of the tau PTMs associated with one or more tau peptide fragments with one or more reference levels for the tau peptide fragments, thereby determining whether a subject has a tauopathy.
Claims21
A method comprising:
- (a) obtaining a first sample from a subject having or suspected of having a tauopathy;
- and
- (b) determining the amount of post translational modification (PTM) associated with a set of tau peptide fragments of a tau protein in the first sample, wherein determining the amount of PTM comprises:
- (i) providing a second sample comprising a labeled tau protein;
- (ii) mixing the first sample and the second sample at an initial mixing ratio of tau protein to labeled tau protein to form a mixture;
- (iii)subjecting the mixture to proteolytic digestion, generating tau peptide fragments and labeled tau peptide fragments;
- (iv)quantifying the abundance of the tau peptide fragments and the labeled tau peptide fragments by performing scheduled selected reaction monitoring acquisition using a selected transition list;
- (v) measuring the ratio of the abundance of the tau peptide fragments and the labeled tau peptide fragments;
- (vi)determining the amount of the tau PTMs associated with the set of tau peptide fragments by comparing the measured ratio for each tau peptide fragment to the initial mixing ratio, wherein the extent of deviation from the initial mixing ratio indicates the amount of PTMs in the tau peptide fragment, wherein the set of tau peptide fragments comprises:
- SEQ ID NO:
- 3, SEQ ID NO:
- 8, SEQ ID NO:
- 12, SEQ ID NO:
- 13, SEQ ID NO:
- 14, SEQ ID NO:
- 16, and SEQ ID NO:
- 17.
A method for quantifying the amount of post-translational modifications on a tau protein from a subject, the method comprising:
- (a) providing a first sample comprising an unlabeled tau protein;
- (b) providing a second sample comprising a labeled tau protein;
- (c) mixing the first sample and the second sample at an initial mixing ratio of unlabeled tau protein to labeled tau protein to form a mixture;
- (d) subjecting the mixture to proteolytic digestion, generating unlabeled tau peptide fragments and labeled tau peptide fragments;
- (e) quantifying the abundance of the unlabeled tau peptide fragments and the labeled tau peptide fragments by performing scheduled selected reaction monitoring acquisition using a selected transition list;
- (f) measuring the ratio of the abundance of the unlabeled tau peptide fragments and the labeled tau peptide fragments, and comparing the ratio for each peptide fragment to the initial mixing ratio, wherein the extent of deviation from the initial mixing ratio indicates the amount of PTMs in the unlabeled protein, wherein the tau peptide fragments comprise:
- SEQ ID NO:
- 3, SEQ ID NO:
- 8, SEQ ID NO:
- 12, SEQ ID NO:
- 13, SEQ ID NO:
- 14, SEQ ID NO:
- 16, and SEQ ID NO:
- 17;
- and
- (g) quantifying the amount of PTMs in the tau protein of the first sample.
A method for quantifying the amount of post-translational modifications on a tau protein, the method comprising:
- (a) providing a mixture comprising a first sample comprising an unlabeled tau protein fragments and a second sample comprising labeled tau peptide fragments;
- (b) quantifying the abundance of the unlabeled tau peptide fragments and the labeled tau peptide fragments by performing scheduled selected reaction monitoring acquisition using a selected transition list;
- (c) measuring the ratio of the abundance of the unlabeled tau peptide fragments and the labeled tau peptide fragments, and comparing the ratio for each peptide fragment to the initial mixing ratio, wherein the extent of deviation from the initial mixing ratio indicates the amount of PTMs in the unlabeled protein, wherein the tau peptide fragments comprise:
- SEQ ID NO:
- 3, SEQ ID NO:
- 8, SEQ ID NO:
- 12, SEQ ID NO:
- 13, SEQ ID NO:
- 14, SEQ ID NO:
- 16, and SEQ ID NO:
- 17;
- and
- (d) quantifying the amount of PTMs in the tau protein of the first sample.
Dependencies are inferred from the claim text.