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Liquid chromatography‐selected reaction monitoring mass spectrometry (LC‐SRM) is not only a proven tool for clinical chemistry, but also a versatile method to enhance the capability to quantify biomarkers for tumor biology research. As the treatment of cancer continues to evolve, the ability to assess multiple biomarkers to assign cancer phenotypes based on the genetic background and the signaling of the individual tumor becomes paramount to our ability to treat the patient. In breast cancer, the American Society of Clinical Oncology (ASCO) has defined biomarkers for patient assessment to guide selection of therapy: estrogen receptor, progesterone receptor, and the HER2/Neu receptor tyrosine kinase; therefore, these proteins were selected for LC‐SRM assay development. Detailed molecular characterization of these proteins is necessary for patient treatment, so expression and phosphorylation assays have been developed and applied. In addition, other LC‐SRM assays were developed to further evaluate tumor biology (e.g. Ki‐67 for proliferation and vimentin for tumor aggressiveness related to the epithelial‐to‐mesenchymal transition). These measurements combined with biomarkers for tissue quality and histological content are implemented in a three tier multiplexed assay platform, which is translated from cell line models into frozen tumor tissues banked from breast cancer patients.
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