Method for diagnosing or monitoring kidney function or diagnosing kidney dysfunction

1. A method for (a) diagnosing or monitoring kidney function in a subject or (b) diagnosing kidney dysfunction in a subject or (c) predicting or monitoring the risk of an adverse event in a subject with kidney disease wherein said adverse event is selected from the group comprising worsening of kidney dysfunction including kidney failure, loss of kidney function and end-stage kidney disease or death due to kidney dysfunction including kidney failure, loss of kidney function and end-stage kidney disease or (d) predicting or monitoring the success of a therapy or intervention in a subject with kidney disease comprising:

determining the level of Pro-Enkephalin or fragments thereof in a bodily fluid obtained from said subject; and

(a) correlating said level of Pro-Enkephalin or fragments thereof with kidney function in a subject, or

(b) correlating said level of Pro-Enkephalin or fragments thereof with kidney dysfunction, wherein an elevated level above a certain threshold is predictive or diagnostic for kidney dysfunction in said subject, or

(c) correlating said level of Pro-Enkephalin or fragments thereof with said risk of an adverse event in a subject with a kidney disease, wherein an elevated level above a certain threshold is predictive for an enhanced risk of said adverse events, or

(d) correlating said level of Pro-Enkephalin or fragments thereof with success of a therapy or intervention in a subject with a kidney disease, wherein a level below a certain threshold is predictive for a success of therapy or intervention,

wherein said Pro-Enkephalin or fragment is one or more of SEQ ID No. 1 or SEQ ID No. 6,

wherein the level of Pro-Enkephalin or fragments thereof is determined by using an assay comprising two binders that bind to two different regions within the region of Pro-Enkephalin that is amino acid 133-140 (LKELLETG, SEQ ID NO. 13) and amino acid 152-159 (SDNEEEVS, SEQ ID No. 14), wherein each of said regions comprises at least 4 amino acids.

2. A method according to claim 1 wherein the binders are antibodies, antibody fragments or a non-Ig-Scaffold binding to Pro-Enkephalin or fragments thereof.

3. A method according to claim 1 comprising

determining the level of immunoreactive analyte by using an assay comprising two binders that bind to two different regions within the region of Pro-Enkephalin that is amino acid 133-140 (LKELLETG, SEQ ID NO. 13) and amino acid 152-159 (SDNEEEVS, SEQ ID No. 14), wherein each of said regions comprises at least 4 amino acids; and

(a) correlating said level of immunoreactive analyte with kidney function in a subject, or

(b) correlating said level of immunoreactive analyte with kidney dysfunction wherein an elevated level above a certain threshold is predictive or diagnostic for kidney dysfunction in said subject, or

(c) correlating said level of immunoreactive analyte with said risk of an adverse event in a subject with a kidney disease, wherein an elevated level above a certain threshold is predictive for an enhanced risk of said adverse events, or

(d) correlating said level of immunoreactive analyte with success of a therapy or intervention in a subject with a kidney disease, wherein a level below a certain threshold is predictive for a success of therapy or intervention,

wherein said Pro-Enkephalin or fragment is one or more of SEQ ID No. 1 or SEQ ID No. 6.

4. A method according to claim 1 , wherein said binders bind to a region within an amino acid sequence which is one or more of SEQ ID No. 1 or 6.

5. A method according to claim 1 , wherein said threshold level of Pro-Enkephalin or fragments thereof is 80 pmol/L.

6. A method according to claim 1 , wherein the level of Pro-Enkephalin is measured with an immunoassay and each of said binders is an antibody, or an antibody fragment or a non-Ig-Scaffold binding to Pro-Enkephalin or fragments thereof.

7. A method according to claim 1 , wherein an assay is used for determining the level of Pro-Enkephalin or fragments thereof and wherein the assay sensitivity of said assay is able to quantify the Pro-Enkephalin or Pro-Enkephalin fragments of healthy subjects and is <15 pmol/L.

8. A method according to claim 1 , wherein said bodily fluid may be selected from the group comprising blood, serum, plasma, urine, cerebrospinal liquid (csf), and saliva.

9. A method according to claim 1 , wherein at least one additional clinical parameter is determined selected from the group comprising: age, BUN, NGAL, Creatinine Clearance, Creatinine and Apache Score.

10. A method according to claim 1 , wherein said determination of Pro-Enkephalin or fragments thereof is performed more than once in one subject.

11. A method according to claim 1 , wherein said monitoring is performed in order to evaluate the response of said subject to preventive and/or therapeutic measures taken.

12. A method according to claim 1 , in order to stratify said subjects into risk groups.

13. A method according to claim 1 , wherein said level of Pro-Enkephalin or fragments thereof are correlated with a risk of death or an adverse event in a subject with a kidney disease, wherein an elevated level above a certain threshold is predictive for an enhanced risk of death or adverse events and wherein said subject with a kidney disease is male.

14. A point-of-care device for performing assays near a subject, wherein said point-of-care device is capable of performing a method of claim 1 and comprises at least two antibodies or antibody fragments directed to amino acid 133-140 (LKELLETG, SEQ ID No. 13) and amino acid 152-159 (SDNEEEVS, SEQ ID NO. 14).

15. A kit which comprises: at least two antibodies or antibody fragments directed to either amino acid 133-140 (LKELLETG, SEQ ID No. 13) or amino acid 152-159 (SDNEEEVS, SEQ ID NO. 14).

16. A method according to claim 1 , wherein said binders do not bind to enkephalin peptides [Met]enkephalin SEQ ID No:3, and [Leu]enkephalin. SEQ ID No: 4.

17. A method according to claim 1 , wherein said binders bind to SEQ ID No. 6.

18. A method according to claim 1 , wherein said binders bind to a region within an amino acid sequence SEQ ID No. 6.

19. A method comprising

determining the level of Pro-Enkephalin fragments in a bodily fluid obtained from a subject using a binder to said Pro-Enkephalin fragments; and

correlating said level of said Pro-Enkephalin fragments with

kidney function in said subject, or

kidney dysfunction in said subject, wherein an elevated level above a certain threshold is predictive or diagnostic for kidney dysfunction in said subject, or

risk of an adverse event in a subject with kidney disease, wherein an elevated level above a certain threshold is predictive for an enhanced risk of said adverse events, or

success of a therapy or intervention in a subject with kidney disease, wherein a level below a certain threshold is predictive for a success of therapy or intervention,

wherein said Pro-Enkephalin fragments is one or more of SEQ ID No. 2, SEQ ID No. 5, SEQ ID No. 6, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10 or SEQ ID No. 11.

20. A method according to claim 19 additionally comprising

determining the level of immunoreactive analyte by using at least one binder that binds to a region within the amino acid sequence of Pro-Enkephalin fragments in a bodily fluid obtained from said subject; and

correlating said level of immunoreactive analyte with

kidney function in said subject, or

kidney dysfunction in said subject, wherein an elevated level above a certain threshold is predictive or diagnostic for kidney dysfunction in said subject, or

risk of an adverse event in said subject with kidney disease, wherein an elevated level above a certain threshold is predictive for an enhanced risk of said adverse events, or

success of a therapy or intervention in a subject with kidney disease, wherein a level below a certain threshold is predictive for a success of therapy or intervention,

wherein said Pro-Enkephalin fragment is one or more of SEQ ID No. 2, SEQ ID No. 5, SEQ ID No. 6, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10 or SEQ ID No. 11.

21. A method for (a) diagnosing or monitoring kidney function in a subject or (b) diagnosing kidney dysfunction in a subject or (c) predicting or monitoring the risk of an adverse events in a subject, wherein said adverse event is selected from the group comprising worsening of kidney dysfunction including kidney failure, loss of kidney function and end-stage kidney disease or death due to kidney dysfunction including kidney failure, loss of kidney function and end-stage kidney disease or (d) predicting or monitoring the success of a therapy or intervention comprising

determining the level of Pro-Enkephalin or fragments thereof in a bodily fluid obtained from said subject; and

(a) correlating said level of Pro-Enkephalin or fragments thereof with kidney function in said subject, or

(b) correlating said level of Pro-Enkephalin or fragments thereof with kidney dysfunction, wherein an elevated level above a certain threshold is predictive or diagnostic for kidney dysfunction in said subject, or

(c) correlating said level of Pro-Enkephalin or fragments thereof with said risk of an adverse event in a subject with kidney disease, wherein an elevated level above a certain threshold is predictive for an enhanced risk of said adverse events, or

(d) correlating said level of Pro-Enkephalin or fragments thereof with success of a therapy or intervention in a subject with kidney disease, wherein a level below a certain threshold is predictive for a success of therapy or intervention,

wherein said Pro-Enkephalin or fragments thereof is one or more of SEQ ID No. 2, SEQ ID No. 5, SEQ ID No. 6, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10 or SEQ ID No. 11,

wherein the level of Pro-Enkephalin or fragments thereof is determined by using an assay comprising a binder, wherein the binder is selected from the group consisting of an antibody, an antibody fragment, or a non-Ig-Scaffold that does not bind to enkephalin peptides [Met]enkephalin SEQ ID No:3.

22. A method according to claim 21 wherein the binder is selected from the group consisting of an antibody, an antibody fragment, or a non-Ig-Scaffold binding to Pro-Enkephalin or fragments thereof that also does not bind to [Lu]enkephalin. SEQ ID No: 4.

23. A method according to claim 21 further comprising

determining the level of immunoreactive analyte in a bodily fluid obtained from said subject by using an assay comprising a binder that binds to a region within the amino acid sequence of Pro-Enkephalin (PENK) or fragments thereof but does not bind to enkephalin peptides [Met]enkephalin SEQ ID No: 3; and

correlating said level of immunoreactive analyte with

kidney function in a subject, or

kidney dysfunction, wherein an elevated level above a certain threshold is predictive or diagnostic for kidney dysfunction in said subject, or

risk of an adverse event in a subject with kidney disease, wherein an elevated level above a certain threshold is predictive for an enhanced risk of said adverse event, or

success of a therapy or intervention in a subject with kidney disease, wherein a level below a certain threshold is predictive for a success of therapy or intervention,

wherein said Pro-Enkephalin or fragment is one or more of SEQ ID No. 2, SEQ ID No. 5, SEQ ID No. 6, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10 or SEQ ID No. 11.

24. A method according to claim 23 wherein the binder is selected from the group consisting of an antibody, an antibody fragment, or a non-Ig-Scaffold binding to Pro-Enkephalin or fragments thereof that also does not bind to [Leu]enkephalin SEQ ID No: 4.

25. A method according to claim 21 , wherein said threshold level of Pro-Enkephalin or fragments thereof is 80 pmol/L.

26. A method according to claim 21 , wherein an assay is used for determining the level of Pro-Enkephalin or fragments thereof and wherein the assay sensitivity of said assay is able to quantify the Pro-Enkephalin or Pro-Enkephalin fragments of healthy subjects and is <15 pmol/L.

27. A point-of-care device for performing assays near a subject, wherein said point-of-care device is capable of performing a method of claim 21 and comprises an antibody or antibody fragment that does not bind to enkephalin peptides [Met]enkephalin SEQ ID No: 3.

28. A kit which comprises:

a) a point-of-care device for performing assays near a subject, capable of performing a method of claim 21 and

b) an antibody or antibody fragment that does not bind to enkephalin peptides [Met]enkephalin SEQ ID No: 3.

29. A method for (a) diagnosing or monitoring kidney function in subject or (b) diagnosing kidney dysfunction in a subject or (c) predicting or monitoring the risk of an adverse events in a subject wherein said adverse event is selected from the group comprising worsening of kidney dysfunction including kidney failure, loss of kidney function and end-stage kidney disease or death due to kidney dysfunction including kidney failure, loss of kidney function and end-stage kidney disease or (d) predicting or monitoring the success of a therapy or intervention comprising

determining the level of Pro-Enkephalin or fragments thereof in a bodily fluid obtained from said subject; and

(a) correlating said level of Pro-Enkephalin or fragments thereof with kidney function in a subject, or

(b) correlating said level of Pro-Enkephalin or fragments thereof with kidney dysfunction, wherein an elevated level above a certain threshold is predictive or diagnostic for kidney dysfunction in said subject, or

(c) correlating said level of Pro-Enkephalin or fragments thereof with said risk of an adverse event in a subject with kidney disease, wherein an elevated level above a certain threshold is predictive for an enhanced risk of said adverse events, or

(d) correlating said level of Pro-Enkephalin or fragments thereof with success of a therapy or intervention in a subject with kidney disease, wherein a level below a certain threshold is predictive for a success of therapy or intervention,

wherein said Pro-Enkephalin or fragment is one or more of SEQ ID No. 2, SEQ ID No. 5, SEQ ID No. 6, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10 or SEQ ID No. 11,

wherein the level of Pro-Enkephalin or fragments thereof is determined by using an assay with an assay sensitivity able to quantify the Pro-Enkephalin or Pro-Enkephalin fragments of healthy subjects and is <15 pmol/L.

30. A method according to claim 29 additionally comprising

determining the level of immunoreactive analyte in a bodily fluid obtained from said subject by using an assay with at least one binder that binds to a region within the amino acid sequence of Pro-Enkephalin (PENK) or fragments thereof with an assay sensitivity able to quantify the Pro-Enkephalin or Pro-Enkephalin fragments of healthy subjects and is <15 pmol/L; and

correlating said level of immunoreactive analyte with:

kidney function in a subject, or

kidney dysfunction, wherein an elevated level above a certain threshold is predictive or diagnostic for kidney dysfunction in said subject, or

risk of an adverse event in a subject with kidney disease, wherein an elevated level above a certain threshold is predictive for an enhanced risk of said adverse event, or

success of a therapy or intervention in a subject with kidney disease, wherein a level below a certain threshold is predictive for a success of therapy or intervention,

wherein said Pro-Enkephalin or fragment is one or more of SEQ ID No. 2, SEQ ID No. 5, SEQ ID No. 6, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10 or SEQ ID No. 11.

31. A method for diagnosing or monitoring kidney function using a kit or assay to detect Pro-Enkephalin or fragments thereof in a subject, wherein said kit or assay comprises at least one antibody capable of binding Pro-Enkephalin or fragments thereof in a sample of bodily fluid obtained from said subject, said method comprising:

determining the level of Pro-Enkephalin or fragments thereof in a sample of bodily fluid obtained from a subject using an assay comprising an antibody that binds to Pro-Enkephalin or fragments thereof;

wherein said Pro-Enkephalin or fragment is one or more of SEQ ID No. 2, SEQ ID No. 5, SEQ ID No. 6, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10 or SEQ ID No. 11

correlating said level of Pro-Enkephalin or fragments thereof with

kidney function in a subject, or

kidney dysfunction wherein an elevated level above a certain threshold is predictive or diagnostic for kidney dysfunction in said subject, or

risk of an adverse event in a subject with kidney disease, wherein an elevated level above a certain threshold is predictive for an enhanced risk of said adverse event, or

success of a therapy or intervention in a subject with kidney disease,

wherein a level below a certain threshold is predictive for a success of therapy or intervention.

32. A method of using a kit or assay according to claim 31 additionally comprising

determining the level of immunoreactive analyte by using an assay comprising an antibody that binds to a region within the amino acid sequence of Pro-Enkephalin or fragments thereof in a bodily fluid obtained from said subject; and

correlating said level of immunoreactive analyte with

kidney function in a subject, or

kidney dysfunction in a subject, wherein an elevated level above a certain threshold is predictive or diagnostic for kidney dysfunction in said subject, or

risk of an adverse event in a subject with kidney disease, wherein an elevated level above a certain threshold is predictive for an enhanced risk of said adverse event, or

success of a therapy or intervention in a subject with kidney disease,

wherein a level below a certain threshold is predictive for a success of therapy or intervention,

wherein said Pro-Enkephalin or fragment is selected from the group consisting of SEQ ID No. 1, SEQ ID No. 2, SEQ ID No. 5, SEQ ID No. 6, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10 and SEQ ID No. 11.

33. A method according to claim 31 , wherein the antibody of said kit or assay does not bind to enkephalin peptides [Me]enkephalin SEQ ID No: 3, and [Leu]enkephalin. SEQ ID No: 4.

34. A method according to claim 31 , wherein the antibody of said kit or assay binds to SEQ ID No. 6.

35. A method according to claim 31 , wherein the antibody of said kit or assay binds to a region within an amino acid sequence selected from the group consisting of SEQ ID No. 1, 2, 5, 6, 7, 8, 9 and 10.

36. A method according to claim 31 , wherein the antibody of said kit or assay binds to a region within an amino acid sequence selected from the group consisting of SEQ ID No. 2, 5, 6, and 10.

37. A method according to claim 31 , wherein the sensitivity of the assay used for determining the level of Pro-Enkephalin or fragments thereof is able to quantify the Pro-Enkephalin or Pro-Enkephalin fragments of healthy subjects and is <15 pmol/L.