	US 11,698,378 B2
	Methods and compositions for tauopathy diagnosis and treatment
Judith AJ Steen, Brighton, MA (US); Hanno Steen, Brighton, MA (US); Waltraud Mair, Cambridge, MA (US); Jan Muntel, Cambridge, MA (US); and Shaojun Tang, Dedham, MA (US)
Assigned to Children's Medical Center Corporation, Boston, MA (US)
Appl. No. 15/762,720
Filed by Judith AJ Steen, Brighton, MA (US); Hanno Steen, Brighton, MA (US); Waltraud Mair, Cambridge, MA (US); Jan Muntel, Cambridge, MA (US); Shaojun Tang, Dedham, MA (US); and Children's Medical Center Corporation, Boston, MA (US)
PCT Filed Sep. 23, 2016, PCT No. PCT/US2016/053357 § 371(c)(1), (2) Date Mar. 23, 2018, PCT Pub. No. WO2017/053739, PCT Pub. Date Mar. 30, 2017.
Claims priority of provisional application 62/232,575, filed on Sep. 25, 2015.
Prior Publication US 2019/0234966 A1, Aug. 1, 2019
Int. Cl. G01N 33/68 (2006.01); C12Q 1/37 (2006.01)

CPC G01N 33/6896 (2013.01) [C12Q 1/37 (2013.01); G01N 2440/00 (2013.01); G01N 2800/2814 (2013.01); G01N 2800/2821 (2013.01)]

21 Claims

1. 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.