Description
Introduction:
Cetuximab is an epidermal growth factor receptor (EGFR) inhibitor used for the treatment of metastatic colorectal cancer, metastatic non-small cell lung cancer and head and neck cancer. Cetuximab is a chimeric (mouse/human) monoclonal antibody given by intravenous infusion that is distributed under the trade name Erbitux in the U.S. and Canada by the drug company Bristol-Myers Squibb and outside the U.S. and Canada by the drug company Merck KGaA. In Japan, Merck KGaA, Bristol-Myers Squibb and Eli Lilly have a codistribution. In July 2009, the FDA approved Cetuximab (Erbitux) for treatment of colon cancer with wild-type KRAS, since it had little or no effect in colorectal tumors harboring a KRAS mutation (this also applied to the EGFR antibody panitumumab).Anti-Drug Antibodies (ADA) may induce unwanted side effects in biopharmaceuticals. Hence, ADA has been subjected to increase in scrutiny by the regulatory authorities using immunogenicity safety studies. ADA has been observed in pre-clinical and clinical studies, resulting in significant changes in toxicology, pharmacokinetics and efficacy. These effects result from the generation of drug-induced (neutralizing) autoantibodies against Cetuximab and can be responsible for allergic reaction, or even anaphylactic shock. This ELISA kit detects Anti-Cetuximab antibodies and may be used for monitoring immunogenicity.
Intended Use:
The KRIBIOLISA Anti-Cetuximab ELISA is used as an analytical tool for quantitative determination of Anti-Cetuximab antibodies in serum, plasma and cell culture supernatant.
Principle:
The method employs the quantitative sandwich enzyme immunoassay technique. Cetuximab is pre-coated onto microwells. Samples and standards are pipetted into microwells and antibodies to Cetuximab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated Cetuximab 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 Anti-Cetuximab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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