Enzyme Immunoassay for the Quantitative Determination of Ovarian Cancer Antigen CA125
in Human Serum

Product Description
Cancer Antigen 125 (CA125) is a surface antigen associated with epithelial ovarian cancer. In serum, CA125 is associated with a high molecular weight glycoprotein. Published studies have indicated that elevated serum CA125 levels can be found in individuals with serious endometroid, clear-cell and undifferentiated ovarian carcinoma.

The serum CA125 concentration is greater than 35 units per ml in 60% of women with ovarian cancer and >80% of patients with disseminated ovarian cancer. The serum CA125 is elevated in 1% of normal healthy women, 3% of normal healthy women with benign ovarian diseases, 6% of patients with non-neoplastic conditions (including but not limited to first trimester pregnancy, menstruation, endometriosis, uterine fibrosis, acute salphingitis, hepatic diseases and inflammation of peritoneum, pericardium or pleura).
Serial determinations of serum CA125 as well as pelvic examination increase the test specificity. Serum CA125 concentration may be useful in monitoring treatment and distinguishing between good response to treatment and progressive malignant disease with poor therapeutic response. To date, CA125 is the most sensitive marker for residual epithelial ovarian cancer. CA125 may also be elevated in patients with lung, cervical, fallopian tube, and uterine cancer and endometriosis.

Principle
The CA125 ELISA test is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay system utilizes a monoclonal antibody directed against a distinct antigenic determinant on the intact CA125 molecule is used for solid phase immobilization (on the microtiter wells).
A rabbit anti-CA125 antibody conjugated to horseradish peroxidase (HRP) is in the antibody-enzyme conjugate solution. The test sample is allowed to react simultaneously with the two antibodies, resulting in the CA125 molecules being sandwiched between the solid phase and enzyme-linked antibodies.
After incubation at 37°C for 90 minutes, the wells are washed with water to remove unbound-labeled antibodies. A solution of TMB Reagent is added and incubated for 20 minutes, resulting in the development of a blue color. The color development is stopped with the addition of Stop Solution changing the color to yellow.
The concentration of CA125 is directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450nm.

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