title: HER-2 assessment in formalin-fixed paraffin-embedded breast cancer tissue by well-based reverse phase protein array.
abstract: BACKGROUND
  The human epidermal growth factor receptor-2 (HER-2) expression level is a critical element for determining the prognosis and management of breast cancer. HER-2 targeted therapy in breast cancer depends on the reliable assessment of HER-2 expression status but current standard methods are lacking a rigorous quantitative assay. To address this challenge, we developed an assessment of HER-2 expression method by well-based reverse phase protein array (RPPA).
  
  
  RESULTS
  Well-based RPPA is based on a robust protein isolation methodology paired with a novel electrochemiluminescence detection system. HER-2 value of well-based RPPA significantly correlated with dot blotting results (R(2) = 0.939). By well-based RPPA, we successfully detected HER-2 expression in 76 human breast formalin-fixed paraffin-embedded tissue samples. We observed 93.4% (71/76) concordance between well-based RPPA and current HER-2 immunohistochemical assessment guideline. When the cutoff level of HER-2 value was set to 0.689 (HER-2/GAPDH) on the basis of receiver-operating characteristic curve, the area under the curve was 0.975 (95% CI, 0.941-1.000). Sensitivity and specificity of well-based RPPA was 92.1% and 94.7%, respectively.
  
  
  CONCLUSIONS
  HER-2 value by well-based RPPA was correlated with the current HER-2 status guideline, suggesting that this normalized HER-2 assessment may offer advantages over unnormalized current immunohistochemical assessment methods.
authors.affiliation: Tissue Array Research Program, Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 USA ; Antibody Characterization Laboratory, Advanced Technology Program, Leidos Biomedical Research, Inc, Frederick, MD USA.
authors.forename: Candice
authors.initials: C
authors.lastname: Perry
authors.affiliation: Tissue Array Research Program, Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 USA.
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Conway
authors.affiliation: Tissue Array Research Program, Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 USA.
authors.forename: Jeong Won
authors.initials: JW
authors.lastname: Ha
authors.affiliation: Institute of Pathology, RWTH Aachen University, Aachen, Germany.
authors.forename: Till
authors.initials: T
authors.lastname: Braunschweig
authors.affiliation: Tissue Array Research Program, Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Morris
authors.affiliation: Tissue Array Research Program, Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 USA.
authors.forename: Kris
authors.initials: K
authors.lastname: Ylaya
authors.affiliation: Tissue Array Research Program, Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 USA ; Department of Obstetrics and Gynecology, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, 425-707 Korea.
authors.forename: Hanbyoul
authors.initials: H
authors.lastname: Cho
authors.affiliation: Tissue Array Research Program, Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 USA.
authors.forename: Joon-Yong
authors.initials: JY
authors.lastname: Chung
authors.affiliation: Tissue Array Research Program, Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 USA.
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Hewitt
keywords: Breast cancer; Formalin-fixed paraffin-embedded; Human epidermal growth factor receptor- 2; Immunohistochemistry; Reverse-phase protein array
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