Diagnosis of cancer by detecting auto-antibodies against vascular endothelial growth factor (VEGF)
a technology of vascular endothelial growth factor and cancer diagnosis, which is applied in the direction of angiogenin, instruments, drug compositions, etc., can solve the problems of uterine fibroids, normal menses, uterine serosa irritation, etc., and achieve the effect of not treating cancer that has spread outside the ovary, not predicting the outcome of cytoreductive surgery, and avoiding uterine serosa irritation
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example 1
[0108]We measured the anti-VEGF autoantibody in serum samples using a sandwich ELISA kit (CellTrend GmbH Luckenwalde, Germany). The microtiter 96-well polystyrene plates were coated with a polypeptide consisting of amino acids 27 to 191 of human VEGF165 of SEQ ID NO:1, derived from cell line SF21. To maintain the conformational epitopes of the protein, 1 mM calcium chloride was added to every buffer. Duplicate samples of a 1:100 serum dilution were incubated at 4° C. for 2 hours. After washing steps, plates were incubated for 60 minutes with a 1:20.000 dilution of horseradish-peroxidase-labeled goat anti-human IgG (Jackson, USA) used for detection. In order to obtain a standard curve, plates were incubated with test sera from an anti-VEGF autoantibody positive index patient. The ELISA was validated according to the FDA's “Guidance for industry: Bioanalytical method validation”.
[0109]VEGF-Auto-Antibodies are not available, a serum sample from a patient with a systemic sclerosis is us...
example 2
[0110]Anti-VEGF antibody levels in serum samples from 132 healthy donors (“Control”) and 201 patients with ovarian cancer (“OvCA”) were measured using the kit and method of example 1. The levels were determined in units / mL. FIG. 1 shows the mean values of the natural logarithm of the VEGF antibody level for OvCA and Control subjects. Patient suffering from ovarian cancer had significantly lower levels (p<0.0001) of anti-VEGF antibodies as compared to healthy controls.
example 3
[0111]Levels of anti-VEGF antibodies were determined in samples from patients suffering from ovarian cancer taken before onset of surgical removal of the cancer. The patients were treated with a platinum analogue after surgical removal of ovarian cancer. Results are shown in FIG. 2. Anti-VEGF antibody levels in samples of patients who survived after the treatment of the cancer were significantly higher than in those who died after said treatment.
[0112]Furthermore, the levels of CA125, an approved marker for ovarian cancer, in the samples were determined and correlated with the anti-VEGF antibody levels. The results are shown in FIG. 3. A significant positive correlation could be observed between CA125 levels an levels of anti-VEGF antibodies (FIG. 3): correlation: 0.296 (p=0.060).
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