US 11,906,530 B2
Methods for the detection of tau protein aggregates
Byron Winslow Caughey, Hamilton, MT (US); Eri Saijo, Hamilton, MT (US); Allison Lindsey Kraus, Hamilton, MT (US); and Michael Anthony Metrick, II, Hamilton, MT (US)
Assigned to The United States of America, as represented by the Secretary Department of Health and Human Services, Bethesda, MD (US)
Appl. No. 16/474,040
Filed by THE UNITED STATES OF AMERICA, as represented by the Secretary, Department of Health and Human Services, Bethesda, MD (US)
PCT Filed Dec. 29, 2017, PCT No. PCT/US2017/069024
§ 371(c)(1), (2) Date Jun. 26, 2019,
PCT Pub. No. WO2018/126180, PCT Pub. Date Jul. 5, 2018.
Claims priority of provisional application 62/440,885, filed on Dec. 30, 2016.
Prior Publication US 2020/0249245 A1, Aug. 6, 2020
Int. Cl. G01N 33/68 (2006.01)
CPC G01N 33/6896 (2013.01) [G01N 2800/2821 (2013.01)] 35 Claims
OG exemplary drawing
 
1. A method of determining whether a subject has a 3R/4R Tauopathy, comprising:
(a) performing a seeded Tau polymerization assay on a biological sample from the subject, comprising:
(i) contacting the biological sample with a purified recombinant truncated Tau protein, wherein the recombinant truncated Tau protein consists of SEQ ID NO: 43 or SEQ ID NO: 44, to form a reaction mixture;
(ii) incubating the reaction mixture to permit coaggregation of disease-associated Tau protein (TD) present in the biological sample with the recombinant truncated Tau protein;
(iii) maintaining incubation conditions that promote coaggregation of the recombinant truncated Tau protein with the TD to result in a conversion of the recombinant truncated Tau protein to amyloid Tau protein aggregates while inhibiting development of spontaneously forming Tau protein aggregates (rT(spon)); and
(iv) agitating amyloid Tau protein aggregates formed during step (iii), wherein the agitating comprises shaking the reaction mixture in a shaking cycle, wherein each shaking cycle comprises a period of rest and a period of shaking; and
(b) detecting amyloid Tau protein present in the reaction mixture, and wherein detection of amyloid Tau protein in the reaction mixture indicates that the subject has the 3R/4R Tauopathy.