Detection of a specific DEK isoform as a urine-based biomarker for bladder cancer
The present invention is directed to a method of detecting a DEK protein isoform 2 in a human urine sample using ELISA. Methods and compositions for detection of DEK isoform 2 in human urine are provided herein. The presence of DEK isoform 2 in urine is shown to correlate with bladder cancer in humans.
1. A method of detecting bladder cancer in a human, comprising the steps of:
(a) obtaining a urine sample collected from a human, said human is suspected of suffering from bladder cancer;
(b) concentrating said urine sample;
(c) immobilizing a first anti-DEK antibody onto a solid surface, said first anti-DEK antibody recognizes amino acids 1-48 or 83-375 of DEK protein consisting of SEQ ID NO: 3;
(d) adding said concentrated urine sample onto said solid surface after step (c) so as to allow said immobilized first anti-DEK antibody to capture DEK protein onto said solid surface;
(e) removing unbound protein;
(f) adding a second anti-DEK antibody, said second anti-DEK antibody recognizes amino acids 1-48 or 83-375 of DEK protein, so as to allow said second anti-DEK antibody to bind to said captured DEK protein to form a complex wherein said first anti-DEK antibody and said second anti-DEK antibody recognize a different region of DEK protein; and
(g) detecting said complex,
wherein the presence of said complex is indicative of occurrence of bladder cancer in said human.
2. The method of claim 1 , wherein said concentrating step is performed by filtration concentration of said urine sample.
3. The method of claim 1 , wherein said concentrated urine sample is at least 20 fold concentrated as compared to neat urine.
4. The method of claim 1 , wherein said concentrated urine sample is at least 30 fold more concentrated as compared to neat urine.
5. The method of claim 1 , wherein said first anti-DEK antibody is a monoclonal antibody or a polyclonal antibody.
6. The method of claim 1 , wherein said second anti-DEK antibody is labeled with horse-radish peroxidase.
7. A method of detecting DEK protein in a urine sample of a human, comprising the steps of:
(a) obtaining a urine sample collected from a human, said human is having bladder cancer;
(b) concentrating said urine sample;
(c) immobilizing a first anti-DEK antibody onto a solid surface, said first anti-DEK antibody recognizes amino acids 1-48 or 83-375 of DEK protein consisting of SEQ ID NO: 3;
(d) adding said concentrated urine sample onto said solid surface after step (c) so as to allow said immobilized first anti-DEK antibody to capture DEK protein present in said urine sample onto said solid surface;
(e) removing unbound protein;
(f) adding a second anti-DEK antibody, said second anti-DEK antibody recognizes amino acids 1-48 or 83-375 of DEK protein, so as to allow said second anti-DEK antibody to bind to said captured DEK protein to form a complex, wherein said first anti-DEK antibody and said second anti-DEK antibody recognize a different region of DEK protein; and
(g) detecting said bound DEK protein with said second anti-DEK antibody complex.
8. The method of claim 7 , wherein said concentrating step is performed by filtration concentration of said urine sample.
9. The method of claim 7 , wherein said concentrated urine sample is at least 20 fold concentrated as compared to neat urine.
10. The method of claim 7 , wherein said concentrated urine sample is at least 30 fold more concentrated as compared to neat urine.
11. The method of claim 7 , wherein said first anti-DEK antibody is a monoclonal antibody or a polyclonal antibody.
12. The method of claim 7 , wherein said second anti-DEK antibody is labeled with horse-radish peroxidase.