Ipilimumab ELISA (YERVOY)

This Ipilimumab ELISA is for determination of Ipilimumab in human serum, plasma and cell culture supernatant.

Assay Background: Ipilimumab, sold by the trade name Yervoy, is used to treat last stage, metastatic melanoma. A monoclonal antibody, Ipilimumab binds to protein receptor CTLA-4. CTLA-4 functions by down regulating immune responses, which prevents destruction of cancer cells. Ipilimumab`s binding to CTLA-4 inhibits its function, boosting the lymphocyte response to the melanoma cells.

Assay Principle: The method employs the quantitative sandwich enzyme immunoassay technique. Antibodies to Ipilimumab are pre-coated onto microwells. Samples and standards are pipetted into microwells and human Ipilimumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated anti- Ipilimumab antibody is pipetted and incubated. After washing microwells in order to remove any non-specific binding, the ready to use substrate solution (TMB) is added to microwells and color develops proportionally to the amount of Ipilimumab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.