Identification of cancer protein biomarkers using proteomic techniques
The claimed invention describes methods to diagnose or aid in the diagnosis of cancer. The claimed methods are based on the identification of biomarkers which are particularly well suited to discriminate between cancer subjects and healthy subjects. These biomarkers were identified using a unique and novel screening method described herein. The biomarkers identified herein can also be used in the prognosis and monitoring of cancer. The invention comprises the use of leptin, prolactin, OPN and IGF-II for diagnosing, prognosis and monitoring of ovarian cancer.
1. A kit for diagnosing cancer comprising reagents that consist essentially of primary monoclonal antibodies or antibody fragments that specifically detect biomarker proteins leptin, prolactin, osteopontin (OPN) and insulin-like growth factor II (IGF-II), and secondary antibodies that bind to the primary monoclonal antibodies or antibody fragments and are conjugated to a detectable label.
2. The kit of claim 1 , further comprising a reference sample or a predetermined standard.
3. The kit of claim 1 , wherein the primary monoclonal antibodies or antibody fragments are attached to a solid support.
4. The kit of claim 1 , further comprising a reagent for detecting the detectable label.
5. The kit of claim 4 , wherein the reagent for detecting the detectable label is an enzyme or a substrate.
6. The kit of claim 1 , further comprising one or more reagents selected from a buffering agent, a preservative, and a protein stabilizing agent.
7. The kit of claim 1 , wherein each reagent of the kit is enclosed within an individual container and all of the containers are enclosed within a single package.
8. The kit of claim 1 , further comprising instructions for interpreting a result of an assay performed using the kit.
9. The kit of claim 1 , wherein the primary monoclonal antibodies or antibody fragments are synthetic.
10. The kit of claim 1 , wherein the secondary antibodies are synthetic.
11. A kit for diagnosing cancer comprising reagents that consist essentially of primary monoclonal antibodies or antibody fragments that specifically detect biomarker proteins leptin, prolactin, osteopontin (OPN), insulin-like growth factor II (IGF-II), and at least one biomarker protein selected from 6Ckine, angiotensin converting enzyme (ACE), brain-derived neurotrophic factor (BDNF), E-Selectin, epidermal growth factor (EGF), eotaxin-2 (Eot-2), epidermal growth factor receptor 1 (ErbB1), follistatin, hemofiltrate CC chemokine 4 (HCC4), herpes virus entry mediator (HVEM), insulin-like growth factor binding protein 2 (IGFBP-2), interleukin-17 (IL-17), interleukin 1 soluble receptor II (IL-1 sRII), interleukin 2 soluble receptor alpha (IL-2 sRα), macrophage colony stimulating factor receptor (M-CSF R), macrophage migration inhibitory factor (MIF), macrophage inflammatory protein 1 alpha (MIP-1α), macrophage inflammatory protein 3 beta (MIP3β), matrix Metalloproteinase-8 (MMP-8), matrix metalloproteinase 7 (MMP-7), myeloid progenitor inhibitory factor 1 (MPIF-1), pulmonary and activation-regulated chemokine (PARC), platelet-derived growth factor receptor beta (PDGF Rβ), protein C, tumor necrosis factor receptor I (TNF-RI), tumor necrosis factor alpha (TNF-α), soluble Vascular Adhesion Protein-1 (sVAP-1), vascular endothelial growth factor receptor 2 (VEGF R2), VEGF receptor 3 (VEGF R3), human stratum corneum chymotryptic enzyme (HSCCE), kallikrein 4, kallikrein 5, kallikrein 6 (protease M), kallikrein 8, kallikrein 9, kallikrein 10, cancer antigen 125 (CA 125), CA15-3, CA19-9, OVX1, lysophosphatidic acid (LPA), carcinoembryonic antigen (CEA), macrophage colony-stimulating factor (M-CSF), prostasin, CA54-61, CA72, HMFG2, interleukin-6 (IL-6), interleukin-10 (IL-10), LSA, NB70K, PLAP, TAG72, TPA, UGTF, WAP four-disulfide core domain 2 (HE4), matrix metalloprotease 2, tetranectin, inhibin, mesothelin, MUC1, vascular endothelial growth factor (VEGF) NOTCH3, E2F transcription factor 3 (E2F3), GTPase activating protein (RACGAP1), hemotological and neurological expressed 1 (HN1), apolipoprotein A1, laminin, claudin 3 (CLDN3), claudin 4, tumor-associated calcium signal transducer 1 (TROP-1/Ep-CAM), tumor-associated calcium signal transducer 2 (TROP-2), ladinin 1, S100A2, SERPIN2 (PAI-2), CD24, lipocalin 2, matriptase (TADG-15), stratifin, transforming growth factor-beta receptor III (TGF-β-RIII), platelet-derived growth factor receptor alpha, SEMACAP3, ras homology gene family member I (ARHI), thrombospondin 2, disabled-2/differentially expressed in ovarian carcinoma 2 (Dab2/DOC2), and haptoglobin-alpha subunit, and secondary antibodies that bind to the primary monoclonal antibodies or antibody fragments and are conjugated to a detectable label.
12. The kit of claim 11 , further comprising a reference sample or a predetermined standard.
13. The kit of claim 11 , wherein the primary monoclonal antibodies or antibody fragments are attached to a solid support.
14. The kit of claim 11 , further comprising a reagent for detecting the detectable label.
15. The kit of claim 14 , wherein the reagent for detecting the detectable label is an enzyme or a substrate.
16. The kit of claim 11 , further comprising one or more reagents selected from a buffering agent, a preservative, and a protein stabilizing agent.
17. The kit of claim 11 , further comprising instructions for interpreting a result of an assay performed using the kit.
18. The kit of claim 11 , wherein the primary monoclonal antibodies or antibody fragments are synthetic.
19. The kit of claim 11 , wherein the secondary antibodies are synthetic.
20. The kit of claim 11 , wherein each reagent of the kit is enclosed within an individual container and all of the containers are enclosed within a single package.
21. A kit for diagnosing cancer comprising reagents that consist essentially of primary monoclonal antibodies or antibody fragments that specifically detect biomarker proteins leptin, prolactin, osteopontin (OPN), insulin-like growth factor II (IGF-II), and cancer antigen 125 (CA 125), and secondary antibodies that bind to the primary monoclonal antibodies or antibody fragments and are conjugated to a detectable label.
22. The kit of claim 21 , further comprising a reference sample or a predetermined standard.
23. The kit of claim 21 , wherein the primary monoclonal antibodies or antibody fragments are attached to a solid support.
24. The kit of claim 21 , further comprising a reagent for detecting the detectable label.
25. The kit of claim 24 , wherein the reagent for detecting the detectable label is an enzyme or a substrate.
26. The kit of claim 21 , further comprising one or more reagents selected from a buffering agent, a preservative, and a protein stabilizing agent.
27. The kit of claim 21 , wherein each reagent of the kit is enclosed within an individual container and all of the containers are enclosed within a single package.
28. The kit of claim 21 , further comprising instructions for interpreting a result of an assay performed using the kit.
29. The kit of claim 21 , wherein the primary monoclonal antibodies or antibody fragments are synthetic.
30. The kit of claim 21 , wherein the secondary antibodies are synthetic.