Description
Introduction:
Trastuzumab is a humanized monoclonal antibody which targets Her-2/neu receptor. It binds to an extracellular domain of this receptor and inhibits HER2 homodimerization, thereby preventing HER2-mediated signaling. It may also upregulate cell cycle inhibitors such as p21 and p27. Trastuzumab is also thought to facilitate antibody-dependent cellular cytotoxicity, leading to the death of cells that express HER2. It is approved for use in treating breast cancers and is being tested for other cancers that overexpress HER2/neu. 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 Trastuzumab and can be responsible for allergic reaction, or even anaphylactic shock. This ELISA kit detects Anti-Trastuzumab antibodies and may be used for monitoring immunogenicity.
Intended Use:
The KRIBIOLISA Anti-Trastuzumab (Herceptin) ELISA is used as an analytical tool for quantitative determination of Anti-Trastuzumab antibodies in human serum and plasma.
Principle:
The method employs the quantitative sandwich enzyme immunoassay technique. Trastuzumab is pre-coated onto microwells. Samples and standards are pipetted into microwells and antibodies to Trastuzumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated Trastuzumab is pipetted and incubated. 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 Anti-Trastuzumab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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