Methods of determining a treatment protocol for and/or a prognosis of a patient's recovery from a brain injury

• US 10,393,759 B2
• Filed: 09/19/2016
• Issued: 08/27/2019
• Est. Priority Date: 04/12/2011
• Status: Active Grant

• Current Assignee: Quanterix Corporation
• Original Assignee: Quanterix Corporation
• Inventors: Wilson, David, Zetterberg, John Henrik, Blennow, Kaj, Randall, Jeffrey D.
• Primary Examiner(s): Weidner, Adam
• Application Number:
  US15/269,142
• Publication Number:
  US 20170160292A1
• Time in Patent Office: 1,072 Days
• Field of Search: None
• CPC Class Codes: G01N 2333/47 Assays involving proteins of known structure or function as defined in the subgroups
• CPC Class Codes: G01N 2333/4709 Amyloid plaque core protein
• CPC Class Codes: G01N 2800/28 Neurological disorders
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/6896 Neurological disorders, e.g. Alzheimer's disease

Abstract

The present invention, in some embodiments, generally relates to methods of determining a treatment protocol for and/or a prognosis of a patient'"'"'s recovery from a brain injury. In some embodiments, the brain injury results from a hypoxic event. In some embodiments, methods are provided for determining a measure of the concentration of tau protein in a patient sample containing or suspected of containing tau protein.

Claims

1. A method of determining a prognosis of a patient'"'"'s recovery from a brain injury comprising:

(a) performing an assay on each of a plurality of blood samples obtained from the patient following the brain injury and/or plasma and/or serum derived from each of the blood samples to determine a measure of the tau protein molecule concentration in each sample,

wherein the plurality of samples are obtained from the patient over a period of time of at least about 48 hours,

wherein at least one of the plurality of samples is obtained within 12 hours of the brain injury,

wherein the assay is characterized by a limit of detection of tau protein molecules that is less than about 0.2 pg/mL; and

comprises;

i. exposing a plurality of capture objects that each include a binding surface having affinity for tau protein molecules to the blood sample;

ii. immobilizing at least a portion of the tau protein molecules with respect to the plurality of capture objects such that at least some of the capture objects associate with at least one tau protein molecule and a statistically significant fraction of the capture objects do not associate with any tau protein molecules;

iii. determining a proportion of the plurality of capture objects that associated with a tau protein molecule in step ii; and

iv. determining a measure of tau protein molecule concentration in the blood sample based at least in part on the proportion determined in step iii;

(b) determining an area under a curve of a graph of the measure of the tau protein molecule concentration in the plurality of samples versus time, wherein the area is determined for the total period of time and/or a second peak in the tau protein molecule concentration; and

(c) determining a prognosis of the patient'"'"'s recovery from the brain injury and/or a method of treatment based on the area under the curve determined in step (b).

2. The method of claim 1, wherein the brain injury results from a hypoxic event.

3. The method of claim 1, wherein the brain injury comprises a hypoxic event caused by cardiac arrest.

4. The method of claim 1, wherein the brain injury comprises a hypoxic event caused by stroke.

5. The method of claim 1, wherein the brain injury comprises a hypoxic event caused by an ischemic event.

6. The method of claim 1, wherein the brain injury comprises a hypoxic event caused by a thrombosis.

7. The method of claim 1, further comprising determining a method of treatment based at least in part on the area under the curve determined in step (b).

8. The method of claim 1, wherein tau protein molecules are spatially segregated into a plurality of locations, wherein a statistically significant fraction of the locations contain a single tau protein molecule and a statistically significant fraction of the locations do not contain any tau protein molecules.

9. The method of claim 8, wherein the plurality of locations comprises a plurality of reaction vessels.

10. The method of claim 8, wherein the plurality of locations comprises plurality of droplets.

11. The method of claim 1, wherein the plurality of capture objects comprises a plurality of beads.

12. The method of claim 1, further comprising plotting the measure of the tau protein molecule concentration versus time on a graph to determine the area under the curve.

13. The method of claim 1, wherein each of the tau protein molecule is selected from the group consisting of tau 23 (352, 0N3R), tau 24 (383, 0N4R), tau 37 (381, 1N3R), tau 34 (412, 1N4R), tau 39 (410, 2N3R), and tau 40 (441, 2N4R).

14. The method of claim 13, wherein at least a portion of the tau protein molecules are phosphorylated.

15. The method of claim 1, wherein the patient'"'"'s prognosis is correlated with a Cerebral Performance Category (CPC) score including classifications of good or poor,

wherein a good CPC score indicates a prognosis of the patient being conscious and alert with normal function or only slight disability or conscious and alert with moderate disability after a selected period of time, and

wherein a poor CPC score indicates a prognosis of the patient being conscious with the patient having a severe disability, being comatose or in a persistent vegetative state, or being brain dead or suffering death from other causes after a selected period of time.

16. The method of claim 15, wherein the prognosis is good for the recovery from the brain injury if the area under the curve is less than about 800.

17. The method of claim 15, wherein the prognosis is poor for the recovery from the brain injury if the area under the curve is greater than about 800.

18. A method for determining a measure of tau protein from a plurality of blood or blood derived samples obtained from a patient following a brain injury, comprising:

(a) performing an assay on each of the plurality of samples obtained from the patient following the brain injury and/or plasma and/or serum derived from each of the samples to determine a measure of the tau protein molecule concentration in each sample,

wherein the plurality of samples are obtained from the patient over a period of time of at least about 48 hours,

wherein at least one of the plurality of samples is obtained within 12 hours of the brain injury,

wherein the assay comprises

i. detecting tau protein molecules in each sample with single molecule resolution such that the limit of detection of tau protein molecules of the assay is less than about 0.2 pg/mL; and

ii. determining the measure of tau protein molecule concentration in each blood sample based on the results of the detecting step; and

(b) determining an area under a curve of a graph of the measure of the tau protein molecule concentration in the plurality of samples versus time, wherein the area is determined for the total period of time and/or a second peak in the tau protein molecule concentration.

19. The method of claim 18, wherein the assay further comprises:

a) exposing a plurality of capture objects that each include a binding surface having affinity for tau protein molecules to the sample;

b) immobilizing at least a portion of the tau protein molecules with respect to the plurality of capture objects such that at least some of the capture objects associate with at least one tau protein molecule and a statistically significant fraction of the capture objects do not associate with any tau protein molecules;

c) determining a proportion of the plurality of capture objects that associated with a tau protein molecule in step b; and

d) determining a measure of tau protein molecule concentration in the blood sample based at least in part on the proportion determined in step c.

20. The method of claim 19, wherein the assay comprises, between step b and step c, spatially segregating at least a portion of the capture objects subjected to the immobilizing step into a plurality of locations.

21. The method of claim 20, further comprising addressing at least a portion of the plurality of locations subjected to the spatially segregating step and determining the number of said locations containing a tau protein molecule.

22. The method of claim 19, wherein the plurality of capture objects comprises a plurality of paramagnetic beads covalently attached to anti-tau antibodies.

23. The method of claim 20, wherein the plurality of locations comprises a plurality of reactions vessels.

24. The method of claim 23, wherein the average volume of the plurality of reaction vessels is between about 10 attoliters and about 100 picoliters.

25. The method of claim 23, wherein the average volume of the plurality of reaction vessels is between about 1 femtoliter and about 1 picoliter.