Cetuximab ELISA (HERCEPTIN)

Intended Use:This Cetuximab ELISA is a rapid and easy method for the quantitative determination of Cetuximab in human serum, plasma and cell culture supernatant.Assay Background:Cetuximab (Erbitux) is an epidermal growth factor receptor (EGFR) inhibitor used for the treatment of metastatic colorectal cancer and head and neck cancer. Cetuximab is a chimeric (mouse/human) monoclonal antibody given by intravenous infusion. In July 2009, the FDA approved Erbitux for treatment of KRAS wild type colon cancer, since Cetuximab had little or no effect in colorectal tumors harboring a KRAS mutation. Assay Principle:The method employs the quantitative sandwich enzyme immunoassay technique. Antibodies to Cetuximab are pre-coated onto microwells. Samples and standards are pipetted into microwells and human Cetuximab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated anti- Cetuximab antibody is pipetted and incubated with samples. After washing microwells in order to remove any nonspecific binding, the ready to use substrate solution (TMB) is added to microwells and color develops proportionally to the amount of Cetuximab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.