IP Library Granted Patent US 9,910,048
Granted Patent B2
US 9,910,048 · App. 14/649,440 · Granted Mar 6, 2018

Method for detection of aggregates in biological samples

Inventors: Marc Diamond (St. Louis, MO); Najla Kfoury (St. Louis, MO); Brandon B. Holmes (St. Louis, MO); Jennifer L. Furman (St. Louis, MO)
Assignee: WASHINGTON UNIVERSITY
G01N33/6896G01N33/5008G01N2333/4703G01N2800/2821G01N2800/2835
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 9,910,048
App. No.
14/649,440
Granted
Mar 6, 2018
Kind
B2
Abstract

The present invention provides a method for detecting protein aggregates in a biological sample, and uses thereof.

Claims (5)

1. A method for detecting one or more tau aggregates in a biological sample, the method comprising the steps of: (a) providing at least one mammalian cell comprising a first protein linked to a first reporter and a second protein linked to a second reporter, wherein the first protein and the second protein each comprise a similar aggregation-prone domain, such that the first protein and the second protein aggregate and produce a detectable signal; (b) contacting the at least one mammalian cell with a biological sample comprising a protein aggregate at a concentration equivalent to about 1 pM to about 100 nM of monomers, the protein aggregate comprising an aggregation-prone domain similar to the first and second protein, such that the protein aggregate is taken up by the cell; (c) measuring a fluorescence resonance energy transfer (FRET) signal by flow cytometry; and (d) comparing the amount of the detectable signal in the biological sample to the amount of a detectable signal produced by a control, wherein a change in the detectable signal indicates one or more protein aggregates are in the biological sample, wherein the mammalian cell is a HEK293T cell, the aggregation-prone domain of the first and second protein comprises a tau repeat domain with a P301S mutation, the first reporter is CFP and the second reporter is YFP.

2. The method of claim 1 , wherein the biological sample is brain lysate.

3. The method of claim 1 , further comprising an additional step before contacting the at least one mammalian cell with the biological sample, the additional step selected from the group consisting of (i) sonicating the biological sample, and (ii) a lipid-based transfection reagent.

4. The method of claim 2 , wherein the biological sample is in contact with the at least one mammalian cell for at least 4 hours.

5. The method of claim 1 , wherein the mammalian cell is a tau RD P301S monoclonal line ATCC CRL-3275.

Assignments (2)
CONFIRMATORY LICENSE Recorded Mar 31, 2017
From: WASHINGTON UNIVERSITY
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 042126/0166 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 17, 2015
From: DIAMOND, MARC; KFOURY, NAJLA; HOLMES, BRANDON; FURMAN, JENNIFER L.
To: WASHINGTON UNIVERSITY
Reel/Frame 036341/0118 →
Continuity (3)
Provisional Application 61732658 · Dec 3, 2012
Provisional Application 61784178 · Mar 14, 2013
Related Publication 20150309054A1 · Oct 29, 2015