Trastuzumab ELISA (HERCEPTIN)

Intended Use: This Trastuzumab ELISA is a rapid and easy method for the quantitative determination of Trastuzumab in human serum, plasma and cell culture supernatant.

Other features include a standard, validated protocol with ready to use components.

Mechanism of action: Trastuzumab is a humanized monoclonal antibody which targets Her-2/neu receptor. Trastuzumab is approved for use in treating breast cancers and is being tested for other cancers that over expresses HER-2/neu. In such cancer cells, the HER2 protein can be expressed up to 100 times more than in normal cells. This overexpression leads to strong and constant proliferative signaling and hence, tumor formation. Overexpression of HER2 also causes deactivation of checkpoints, allowing for even greater increases in proliferation. Trastuzumab binds to domain IV of the extracellular segment of the HER2/neu receptor. Monoclonal antibodies that bind to this region have been shown to reverse the phenotype of HER2/neu expressing tumor cells.

Assay Principle: The method employs the quantitative sandwich enzyme immunoassay technique. Antibodies to Trastuzumab are pre-coated onto microwells. Samples and standards are pipetted into microwells and human Trastuzumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated anti-Trastuzumab 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 Trastuzumab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.