Biomarker for detecting cancer
Methods for detecting colorectal cancer in a subject by detection of a galactose-containing 40 kDa molecule in a serum sample from the subject. Methods for quantifying the amount of a galactose-containing molecule in a serum sample are also provided.
1. A method for detection or diagnosis of colorectal cancer in a subject, comprising:
(a) obtaining a serum sample of the subject;
(b) desialylating the serum sample;
(c) contacting the desialylated serum sample with an antibody that binds haptoglobin to form a complex;
(d) quantitating the complex with a detectable lectin that binds galactose in order to detect or diagnose colorectal cancer using a quantitative immune-detection assay, wherein the presence of a galactose-containing haptoglobin glycoform is associated with the presence of colorectal cancer, wherein the quantitative immune-detection assay is not a Western blot, and wherein the detectable lectin that binds galactose is selected from the group consisting of Ricinus communis lectin, Sophora japonica lectin, Datura stramonium lectin, Erythrina cristagalli lectin, and Lycopersicon esculentum lectin; and
(e) administering an anticancer therapy to the subject having colorectal cancer.
2. The method of claim 1 , wherein step (a) comprises diluting the serum between 25-fold and 50-fold.
3. The method of claim 1 , wherein the quantitative immune-detection assay is an enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunoradiometric assay, fluoroimmunoassay, chemiluminescent assay or bioluminescent assay.
4. The method of claim 3 , wherein the quantitative immune-detection assay is an ELISA.
5. The method of claim 1 , wherein desialylating the serum comprises treating the serum with a mild acid.
6. The method of claim 5 , wherein the mild acid is H 2 SO 4 .
7. The method of claim 1 , wherein desialylating the serum comprises treating the serum with a neuraminidase.
8. The method of claim 1 , wherein the antibody that binds haptoglobin is a polyclonal antibody.
9. The method of claim 1 , wherein the antibody that binds haptoglobin is bound to a substrate or a magnetic bead.
10. The method of claim 8 , wherein the antibody that binds haptoglobin is a polyclonal antibody raised against purified human haptoglobin.
11. The method of claim 1 , further comprising contacting the complex with a wash solution prior to step (d).
12. The method of claim 11 , wherein the wash solution comprises a detergent.
13. The method of claim 1 , wherein quantitating the complex with a detectable lectin, comprises contacting the complex with the detectable lectin and washing the resulting complex with a wash solution prior to said quantitating.
14. The method of claim 1 , wherein quantitating the complex with a detectable lectin comprises detecting an enzymatic activity.
15. The method of claim 14 , wherein the lectin is biotinylated or comprises a conjugated enzyme.
16. The method of claim 15 , wherein quantitating the complex comprises contacting the complex with an avidin reporter conjugate.
17. The method of claim 16 , wherein the reporter is an enzyme.
18. The method of claim 1 , wherein the detectable lectin is Erythrina cristagalli lectin.
19. The method of claim 4 , wherein the quantitative immune-detection assay is a sandwich ELISA.
20. The method of claim 1 , wherein the quantitative immune-detection assay is an ELISA and wherein the detectable lectin is Erythrina cristagalli lectin.
21. The method of claim 1 , further comprising obtaining the level of carcinoembryonic antigen (CEA) in a serum sample from the subject.
22. The method of claim 1 , further comprising obtaining the level of carcinoembryonic antigen (CEA) and galectin-3 ligand in a serum sample from the subject.
23. A method of treating a subject having colorectal cancer comprising administering an anticancer therapy to the subject having colorectal cancer, said subject having been determined to have a colorectal cancer by a method comprising:
(b) desialylating the serum sample from the subject;
(c) contacting the desialylated serum sample with an antibody that binds haptoglobin to form a complex; and
(d) quantitating the complex with a detectable lectin that binds galactose in order to detect or diagnose colorectal cancer using a quantitative immune-detection assay, wherein the presence of a galactose-containing haptoglobin glycoform is associated with the presence of colorectal cancer, wherein the quantitative immune-detection assay is not a Western blot, and wherein the detectable lectin that binds galactose is selected from the group consisting of Ricinus communis lectin, Sophora japonica lectin, Datura stramonium lectin, Erythrina cristagalli lectin, and Lycopersicon esculentum lectin.