Method for the early detection of renal injury

• US 20050272101A1
• Filed: 03/31/2005
• Published: 12/08/2005
• Est. Priority Date: 06/07/2004
• Status: Abandoned Application

• Current Assignee: Children's Hospital & Medical Center, National Institutes of Health (NIH) (Government of the United States of America)
• Original Assignee: National Institutes of Health (NIH) (Government of the United States of America)
• Inventors: Barasch, Jonathan M., Devarajan, Prasad
• Application Number:
  US11/096,113
• Publication Number:
  US 20050272101A1
• Time in Patent Office: Days
• US Class Current: 435/7.900
• CPC Class Codes: G01N 2333/82 Translation products from oncogenes
• CPC Class Codes: G01N 2800/347 Renal failures; Glomerular diseases; Tubulointerstitial diseases, e.g. nephritic syndrome, glomerulonephritis; Renovascular diseases, e.g. renal artery occlusion, nephropathy
• 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 33/573 for enzymes or isoenzymes
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

A method and kit for detecting the immediate or early onset of renal disease and injury, including renal tubular cell injury, utilizing NGAL as an immediate or early on-set biomarker in a sample of blood serum. NGAL is a small secreted polypeptide that is protease resistant and consequently readily detected in the blood serum following renal tubule cell injury. NGAL protein expression is detected predominantly in proximal tubule cells, in a punctuate cytoplasmic distribution reminiscent of a secreted protein. The appearance NGAL in the serum is related to the dose and duration of renal ischemia and nephrotoxemia, and is diagnostic of renal tubule cell injury and renal failure. NGAL detection is also a useful marker for monitoring the nephrotoxic side effects of drugs or other therapeutic agents.

Claims

5. The method for monitoring the effectiveness of a treatment for renal tubular cell injury, comprising the steps of:

a) obtaining a blood serum sample from a mammalian subject experiencing renal tubular cell injury;

b) determining from the serum sample the level of a biomarker selected from an immediate renal tubular cell injury biomarker, an early on-set renal tubular cell injury biomarker, and mixtures thereof, and

c) evaluating the renal tubular cell injury status of the subject.

6. The method according to claim 5 wherein the biomarker comprises NGAL.

7. The method according to claim 5, further comprising the step of obtaining one or more subsequent post-treatment serum samples, and re-evaluating the renal tubular cell injury status of the subject.

8. The method according to claim 7, further comprising the step of continuing the treatment until the presence of the biomarker in the subsequent post-treatment serum samples is reduced to a level wherein the renal tubular cell injury is ameliorated.

9. The method according to claim 5 wherein the step of detecting comprises the steps of:

i) contacting the serum sample with a capture antibody for the biomarker to allow formation of a complex of the antibody and the biomarker; and

ii) detecting the antibody-biomarker complex.

10. The method according to claim 9 wherein the step of detecting the antibody-biomarker complex comprises the steps of:

a) separating any unbound material of the serum sample from the capture antibody-biomarker complex;

b) contacting the capture antibody-biomarker complex with a second antibody for detecting the biomarker, to allow formation of a complex between the biomarker and the second antibody;

c) separating any unbound second antibody from the biomarker-second antibody complex; and

d) detecting the second antibody of the biomarker-second antibody complex.

11. A kit for use in detecting the presence of an immediate or early onset biomarker for renal tubular cell injury in the serum of a subject, comprising:

a) a means for acquiring a quantity of a blood serum sample;

b) a media having affixed thereto a capture antibody capable of complexing with a renal tubular cell injury biomarker selected from an immediate renal tubular cell injury biomarker, an early on-set renal tubular cell injury biomarker, and mixtures thereof; and

c) an assay for the detection of a complex of the renal tubular cell injury biomarker and the capture antibody.

1. A method for the immediate or early on-set detection of a renal tubular cell injury in a mammalian subject, comprising the steps of:

a) obtaining a blood serum sample from a mammalian subject;

b) determining from the serum sample the level of a biomarker selected from an immediate renal tubular cell injury biomarker, an early on-set renal tubular cell injury biomarker, and mixtures thereof, and

c) evaluating the renal tubular cell injury status of the subject.

2. The method according to claim 1 wherein the step of determining the level of the biomarker comprises the steps of:

a) contacting the serum sample with an antibody for the biomarker to allow formation of a complex of the antibody and the biomarker; and

b) detecting the antibody-biomarker complex.

3. The method according to claim 1 wherein the biomarker is NGAL.

4. The method according to claim 3 wherein the mammalian subject is a human patient.

12. The kit according to claim 11 wherein the quantity of the serum sample is less than 1 ml.

13. The kit according to claim 12 wherein the quantity is less than 10 microliters.

14. The kit according to claim 11 wherein the acquiring means comprises an implement comprising a surface, the surface comprising the media.

15. The kit according to claim 11 wherein the acquiring means comprises a container for accepting the serum sample, wherein the serum -contacting surface of the container comprises the media.

16. The kit according to claim 11 wherein the assay comprises an ELISA.

17. The kit according to claim 11 that is a point-of-care kit.

18. The kit according to claim 12 that is a point-of-cave kit.

19. The point-of-care kit according to claim 18 wherein the acquiring means comprises an implement comprising a dip-stick, the dip-stick surface comprising the media.

20. The point-of-care kit according to claim 19 wherein the assay comprises a calorimetric dip-stick assay.

20-1. The method according to claim 17 wherein the biomarker comprises NGAL.