title: Image cytometric validation of breast carcinoma markers (ER, HER2 and MIB-1) using tissue microarrays: rabbit monoclonal vs. FDA-approved antibodies.
abstract: OBJECTIVE
  To use the ACIS III (Dako, Carpinteria, California, U.S.A.) with tissue microarrays (TMAs) to compare rabbit monoclonal antibodies (RMab) for ER, HER2, and MIB-1 with FDA-approved monoclonal (FMab) and polyclonal (FPab) antibodies.
  
  
  METHODS
  TMAs of 43 breast cancers were used. Immunohistochemistry was performed using RMab (LabVision, Fremont, California, U.S.A.): ER (SP1; 1/100), HER2 (SP3; 1/100), and MIB-1 (SP6; 1/200). FMPab (Dako) used: ER (1D5; 1/50), HercepTest kit and MIB-1 (MIB-1; 1/160). The stained TMAs were quantitated visually and by image cytometry (ACIS III).
  
  
  RESULTS
  The overall agreement between RMab and FMab for ER using visual (98.45%) and image analysis (97.56%) was excellent, with a kappa level of 0.89 and 0.94, respectively. For HER2, the overall agreement between RMab and FPab was fair for visual (67.44%) and substantial (87.50%) for image analysis, with a kappa level of 0.32 and 0.72, respectively. For MIB-1, there was fair (64.29%) to poor (43.33%) agreement between MIB-1 RMab and FMab using visual and image analysis, with a kappa level of 0.47 and 0.16, respectively.
  
  
  CONCLUSIONS
  RMabs for ER (SP1) and HER2 (SP3) are almost comparable to their counterpart, FDA antibodies; however, MIB-1 RMab (SP6) shows poor concordance with FMab in TMA. Image analysis shows a better concordance than visual quantitation assessment specifically for ER and HER2.
authors: Department of Pathology, Mayo Clinic, Rochester, Minnesota 55902, USA. nassar.aziza@mayo.edu
authors: Aziza
authors: A
authors: Nassar
authors: Christine M
authors: CM
authors: Norton
authors: Diane
authors: D
authors: Lawson
authors: Cynthia
authors: C
authors: Cohen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor / Q000276:immunology / Q000378:metabolism*; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology*; D005260:Female; D006801:Humans; D019044:Image Cytometry / Q000379:methods / Q000592:standards*; D007150:Immunohistochemistry / Q000379:methods / Q000592:standards*; D011817:Rabbits; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology / Q000378:metabolism; D011960:Receptors, Estrogen / Q000276:immunology / Q000378:metabolism; D015203:Reproducibility of Results; D044767:Ubiquitin-Protein Ligases / Q000276:immunology / Q000378:metabolism; D014481:United States; D014486:United States Food and Drug Administration
chemical_list: D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D018719:Erb-b2 Receptor Tyrosine Kinases; D011960:Receptors, Estrogen; D044767:Ubiquitin-Protein Ligases; C513496:MIB1 ligase, human; C508053:ERBB2 protein, human