Methods, devices and kits for detecting or monitoring acute kidney injury
Methods for detecting acute kidney injury in an individual comprise (a) contacting a body fluid sample from the individual with an assay device including neutrophil gelatinase-associated lipocalin (NGAL) antibody and a detectable label, to allow complexing of NGAL protein in the sample with NGAL antibody, and determining an amount of complex formed between NGAL protein from the sample and NGAL antibody in the assay device using the detectable label, wherein NGAL antibody in the device has binding capacity with more than two NGAL protein epitopes, and wherein the amount of the formed complex represents a level of acute kidney injury. Methods for determining an origin of NGAL protein in a sample from an individual include the step of determining relative amounts of monomeric, dimeric and heterodimeric forms of NGAL protein in the sample and allow improved diagnosis and therefore better targeted treatment.
1. A method for detecting acute kidney injury in a cardiac surgery patient, comprising
(a) detecting an amount of homodimeric NGAL protein in urine samples from the patient prior to and at least one day after cardiac surgery by contacting a respective urine sample from the patient with a capturing antibody which binds monomeric, homodimeric and heterodimeric NGAL proteins and which is immobilized on a first enzyme-linked immunosorbent assay (ELISA) device to allow binding of NGAL proteins in the sample, and detecting bound homodimeric NGAL protein using a first detectable monoclonal antibody which specifically binds homodimeric NGAL protein,
(b) detecting an amount of monomeric NGAL protein in the urine samples from the patient prior to and at least one day after cardiac surgery by contacting the respective urine sample from the patient with a capturing antibody which binds monomeric, homodimeric and heterodimeric NGAL proteins and which is immobilized on a second ELISA device to allow binding of NGAL proteins in the sample, and detecting bound monomeric NGAL protein using a second detectable monoclonal antibody which binds monomeric NGAL protein, and
(c) comparing a first ratio of the detected amount of monomeric NGAL protein to homodimeric NGAL protein in the sample prior to cardiac surgery with a second ratio of the amount of detected monomeric NGAL protein to homodimeric NGAL protein in the sample at least one day after cardiac surgery,
wherein, the second ratio being greater than the first ratio indicates the patient has suffered acute kidney injury and the second ratio being less than the first ratio indicates the patient has not suffered acute kidney injury.
2. The method according to claim 1 , wherein the antibodies are produced from purified NGAL protein.
3. The method according to claim 1 , wherein the capturing antibody which binds monomeric, homodimeric and heterodimeric NGAL proteins in step (a) is a monoclonal antibody.
4. The method according to claim 1 , wherein the capturing antibody which binds monomeric, homodimeric and heterodimeric NGAL proteins in step (b) is a monoclonal antibody.
5. A method for detecting NGAL protein originating from kidney in a cardiac surgery patient urine sample, comprising
(a) providing urine samples from the patient prior to and at least one day after cardiac surgery,
(b) detecting an amount of homodimeric NGAL protein in the urine samples from the patient prior to and at least one day after cardiac surgery by contacting a respective urine sample from the patient with a capturing antibody which binds monomeric, homodimeric and heterodimeric NGAL proteins and which is immobilized on a first enzyme-linked immunosorbent assay (ELISA) device to allow binding of NGAL proteins in the sample, and detecting bound homodimeric NGAL protein using a first detectable monoclonal antibody which specifically binds homodimeric NGAL protein, and
(c) detecting an amount of monomeric NGAL protein in the urine samples from the patient prior to and at least one day after cardiac surgery by contacting the respective urine sample from the patient with a capturing antibody which binds monomeric, homodimeric and heterodimeric NGAL proteins and which is immobilized on a second ELISA device to allow binding of NGAL proteins in the sample, and detecting bound monomeric NGAL protein using a second detectable monoclonal antibody which binds monomeric NGAL protein,
wherein an increase of the ratio of the detected amount of monomeric NGAL protein to homodimeric NGAL protein in the sample at least one day after cardiac surgery over the ratio of the amount of detected monomeric NGAL protein to homodimeric NGAL protein in the sample prior to cardiac surgery indicates the NGAL protein origin is from kidney.
6. The method according to claim 5 , wherein the antibodies are produced from purified NGAL protein.
7. The method according to claim 5 , wherein the capturing antibody which binds monomeric, homodimeric and heterodimeric NGAL proteins in step (b) is a monoclonal antibody.
8. The method according to claim 5 , wherein the capturing antibody which binds monomeric, homodimeric and heterodimeric NGAL proteins in step (c) is a monoclonal antibody.
9. A method for detecting NGAL protein in a cardiac surgery patient urine sample, comprising
(a) providing urine samples from the patient prior to and at least one day after cardiac surgery,
(b) detecting an amount of homodimeric NGAL protein in the urine samples from the patient prior to and at least one day after cardiac surgery by contacting a respective urine sample from the patient with a capturing antibody which binds monomeric, homodimeric and heterodimeric NGAL proteins and which is immobilized on a first enzyme-linked immunosorbent assay (ELISA) device to allow binding of NGAL proteins in the sample, and detecting bound homodimeric NGAL protein using a first detectable monoclonal antibody which specifically binds homodimeric NGAL protein, and
(c) detecting an amount of monomeric NGAL protein in the urine samples from the patient prior to and at least one day after cardiac surgery by contacting the respective urine sample from the patient with a capturing antibody which binds monomeric, homodimeric and heterodimeric NGAL proteins and which is immobilized on a second ELISA device to allow binding of NGAL proteins in the sample, and detecting bound monomeric NGAL protein using a second detectable monoclonal antibody which binds monomeric NGAL protein.
10. The method according to claim 9 , wherein the antibodies are produced from purified NGAL protein.
11. The method according to claim 9 , wherein the capturing antibody which binds monomeric, homodimeric and heterodimeric NGAL proteins in step (b) is a monoclonal antibody.
12. The method according to claim 9 , wherein the capturing antibody which binds monomeric, homodimeric and heterodimeric NGAL proteins in step (c) is a monoclonal antibody.