Description
The Eagle Biosciences Humanized Anti-HER-2 (Herceptin/Trastuzumab) ELISA Assay kit utilizes the “sandwich” technique with HER2 coated to solid phase microtiter plate wells and an antibody to human IgG which is labeled with horseradish peroxidase used for detection. Assay calibrators, controls and diluted patient samples are added directly to wells of a micro titer plate that is coated with HER2 recombinant protein. Subsequently, horseradish peroxidase (HRP) conjugated human IgG antibody is added to each well. After the first incubation period, a “sandwich” of “HER2 - Herceptin – HRP conjugated human IgG antibody” is formed on the surface of the plate wells. The unbound proteins are removed in the subsequent washing step. For the detection of this immunocomplex, the wells are then incubated with a substrate solution in a timed reaction and then measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to the wall of each micro titer well is directly proportional to the amount of Herceptin in the calibrators. A calibration curve is generated by plotting the absorbance versus the respective Herceptin concentration for each calibrator using point-to-point or 4 parameter curve fitting. The concentration of Herceptin in test samples is determined directly from this calibration curve