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Anal Quant Cytol Histol. 2010 Aug;32(4):192-200.

Image cytometric validation of breast carcinoma markers (ER, HER2 and MIB-1) using tissue microarrays: rabbit monoclonal vs. FDA-approved antibodies.

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Department of Pathology, Mayo Clinic, Rochester, Minnesota 55902, USA.



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.


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).


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.


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.

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