Improved assays, cartridges, and kits for the detection of biomarkers, including brain injury biomarkers.

Abstract

This specification discloses improved assays, cartridges, kits, and methods of use thereof for detecting one or more biomarkers of acquired brain injury (ABI), such as traumatic brain injury (TBI), which are not limited to the most frequently cited biomarkers. The improved assays described herein may aid in the diagnosis and assessment of subjects (e.g., human subjects) who have suffered or may have suffered head injury (e.g., TBI) by detecting levels of biomarkers, such as UCH-L1, GFAP, or a combination thereof, in samples taken from subjects (e.g., human subjects).

Claims

1. A cartridge for use in a point-of-care device, including a housing, wherein the housing comprises a pretreatment reagent for use in performing at least one assay, and the pretreatment reagent further comprises at least one nuclease.

2. The cartridge according to claim 1, wherein the housing further comprises a biosensor chip, a sample chamber, and a waste liquid container.

3. The cartridge according to claim 2, wherein the pretreatment reagent includes a printed pretreatment layer in the sample chamber.

4. The cartridge according to claim 3, wherein the pretreatment layer contains about 1.5 U / mL of nuclease.

5. The cartridge according to claim 4, wherein the nuclease is Benzonase® nuclease.

6. The cartridge according to claim 4 or 5, wherein the pretreatment reagent dissolves when it comes into contact with a biological sample containing a biomarker or the target analyte.

7. The cartridge according to claim 6, wherein the amount of nuclease dissolved in the biological sample is approximately 1.0 U / mL.

8. The cartridge according to claim 6 or claim 7, wherein the biomarker or target analyte is GFAP, UCH-L1, or GFAP and UCH-L1.

9. A method for measuring the level of a biomarker in a sample, comprising the following steps: a) The steps of contacting the sample obtained from the subject with a solution containing (1) one or more capture antibodies that bind to an epitope on the biomarker to form a capture antibody-biomarker complex, (2) one or more detection antibodies that bind to an epitope on the biomarker that has not been bound by the capture antibody to form a capture antibody-biomarker-detection antibody complex and include a detectable label, and (3) at least one nuclease, and b) A step of measuring the amount or concentration of the biomarker in the sample based on the signal generated by the detectable label in the capture antibody-biomarker-detection antibody complex. The step includes performing at least one assay that includes, The biomarker is glial fibrillary acidic protein (GFAP), ubiquitin carboxy-terminal hydrolase L1 (UCH-L1), or GFAP and UCH-L1, and the step of contacting the sample with the solution containing at least one nuclease reduces (i) nonspecific conjugate binding during the assay, (ii) aggregate formation during the assay, or (iii) a combination of (i) and (ii). method.

10. a) Before contacting the sample with the capture antibody or the detection antibody; b) After contacting the sample with the capture antibody; or c) After contacting the sample with the capture antibody and the detection antibody The method according to claim 9, comprising the step of contacting the sample with the solution containing at least one nuclease.

11. The method according to claim 9 or claim 10, wherein the biomarker is a TBI biomarker.

12. The method according to any one of claims 9 to 11, wherein the at least one nuclease is an endonuclease.

13. The method according to any one of claims 9 to 12, wherein the at least one nuclease cleaves DNA, RNA, or DNA and RNA.

14. The method according to any one of claims 9 to 13, wherein the solution containing the at least one nuclease contains at least about 1 U / mL of the nuclease.

15. The method according to any one of claims 9 to 14, wherein the solution containing the at least one nuclease contains the nuclease in an amount of about 1 U / mL to about 1.5 U / mL.

16. The method according to any one of claims 9 to 15, wherein the solution containing at least one nuclease does not contain heparin.

17. The method according to any one of claims 9 to 16, wherein the at least one assay is performed using a point-of-care device.

18. The method according to any one of claims 9 to 17, wherein the sample is a whole blood sample.

19. An improvement to an assay for measuring the amount of a biomarker for traumatic brain injury (TBI) in a sample obtained from a subject, comprising using a solution containing at least one nuclease when performing the assay, further wherein the TBI biomarker is glial fibrillary acidic protein (GFAP), ubiquitin carboxy-terminal hydrolase L1 (UCH-L1), or GFAP and UCH-L1.

20. A kit comprising a capture antibody, a detection antibody, and a solution comprising at least one nuclease, wherein the use of the kit for an assay to detect one or more biomarkers reduces (i) nonspecific conjugate binding in the assay, (ii) aggregation formation in the assay, or (iii) a combination of (i) and (iii) compared to an assay performed using a kit that does not comprise at least one nuclease, and further wherein the biomarker is glial fibrillary acidic protein (GFAP), ubiquitin carboxy-terminal hydrolase L1 (UCH-L1), or GFAP and UCH-L1.

21. The kit according to claim 20, wherein the solution containing at least one nuclease does not contain heparin.

Images