Anti-Trastuzumab (HERCEPTIN, HERCLON) ELISA Assay Kit
The Anti-Trastuzumab (HERCEPTIN, HERCLON) ELISA Assay Kit is For Research Use Only
Size: 12×8 wells
Sensitivity: +/-
Incubation Time: 2 hours 20 minutes
Sample Type: Serum, Plasma
Sample Size: 20 µL
Alternative Name: Herceptin, Herclon
Assay Background
Trastuzumab is a recombinant humanized IgGl monoclonal antibody against the HER-2 receptor, a member of the epidermal growth factor receptors which is a photo-oncogene. Over-expressed in breast tumor cells, HER-2 overamplifies the signal provided by other receptors of the HER family by forming heterodimers. The HER-2 receptor is a transmembrane tyrosine kinase receptor that consists of an extracellular ligand-binding domain, a transmembrane region, and an intracellular or cytoplasmic tyrosine kinase domain. It is activated by the formation of homodimers or heterodimers with other EGFR proteins, leading to dimerization and autophosphorylation and/or transphosphorylation of specific tyrosine residues in EGFR intracellular domains. Further downstream molecular signaling cascades are activated, such as the Ras/Raf/mitogen-activated protein kinase {MAPK), the phosphoinositide 3-kinase/ Akt, and the phospholipase Cy (PLCy)/protein kinase C {PKC) pathways that promote cell growth and survival and cell cycle progression. Due to upregulation of HER-2 in tumor cells, hyperactivation of these signaling pathways and abnormal cell proliferation is observed. Trastuzumab binds to the extracellular ligand- binding domain and blocks the cleavage of the extracellular domain of HER-2 to induce its antibody-induced receptor downmodulation, and subsequently inhibits HER-2-mediated intracellular signaling cascades. Inhibition of MAPK and Pl3K/Akt pathways lead to an increase in cell cycle arrest, and the suppression of cell growth and proliferation. Trastuzumab also mediates the activation of antibody-dependent cell-mediated cytotoxicity (ADCC) by attracting the immune cells, such as natural killer (NK) cells, to tumor sites that overexpress HER-2. Intrinsic trastuzumab resistance has been noted for some patients with HER-2 positive breast cancer. Mechanisms involving trastuzumab resistance include deficiency of phosphatase and tensin homologue and activation of phosphoinositide 3- kinase, and the overexpression of other surface receptors, such as insulin-like growth factor.
Assay Principle
Solid phase enzyme-linked immunosorbent assay (ELISA) based on the sandwich principle. Controls and samples (serum or plasma) are incubated in the microtiter plate coated with the drug trastuzumab. After incubation, the wells are washed. Then, horse radish peroxidase (HRP) conjugated probe is added and binds to trastuzumab antibodies captured by the drug trastuzumab on the surface of the wells. Following incubation wells are washed and the bound enzymatic activity is detected by addition of chromogen-substrate. Finally, the reaction is terminated with an acidic stop solution. The color developed is proportional to the amount of trastuzumab antibodies in the sample or controls. The results can be evaluated with using cut-off value.
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