INTRODUCTION

Trastuzumab (Herceptin® ) is a humanized recombinant monoclonal antibody used for the treatment of primary breast cancers overexpressing human epidermal growth factor 2 (HER2). Repeated dosing of Trastuzumab has been reported to induce a neutralizing antibody response resulting in reduce efficacy of Trastuzumab. This kit allows
researchers to qualitatively monitor the presence of anti-Trastuzumab antibodies in biological matrices.

PRINCIPLE OF THE ASSAY

This immunogenicity assay employs the bridging ELISA technique. Capture antibody is precoated onto a 96 well microplate. Quality control and test samples are pipetted into the appropriate wells. Anti- Trastuzumab present in biological matrices is bound by the immobilized capture antibody. After washing away any unbound substances, secondary antibody is added to the wells and after a final wash a detection reagent is added. The plate is washed to remove any unbound antibody-enzyme reagent and a substrate solution is added to the wells for color development.
The color development is proportional to the amount of anti-Trastuzumab present in test samples. Three levels of QC samples give a qualitative reference signal which can be used to determine the level of anti-Trastuzumab antibody in the unknown samples. The color development is stopped and the intensity of the color is measured.