Abstract P1-01-16: Performance evaluation of two ready-to-use antibodies under development for the Dako Omnis automated staining platform on breast carcinoma specimens: Anti-estrogen receptor α clone EP1 and anti-progesterone receptor clone PgR 1294
Authors: Viale G , Dell'Orto P , Falzon M , Fält A , Hicks D , Hoff K , Jakobsen K , Jensen LB , Levy YY , McMahon L , Miller K , Russo L 
Published: May 2016

Abstract
Background: The expression of estrogen receptor alpha (ERα) and progesterone receptor (PR) in breast carcinomas is a strong predictor of the efficacy of hormonal therapy for breast cancer patients as well as providing a degree of prognostic information. Anti-ERα (clone EP1) and anti-PR (clone PgR 1294) configured as FLEX ready-to-use antibodies have been tested on the Dako Omnis automated staining platform. These products are in performance evaluation and are not commercially available. A series of concordance studies were performed to evaluate the performance characteristics of these monoclonal antibodies on breast cancer tissue specimens: anti-ERα clone EP1/Dako Omnis was compared to (a) anti-ERα clone EP1/Autostainer Link 48 (238 specimens) and to (b) anti-ERα clone SP1/Autostainer (116 specimens), and anti-PR clone PgR 1294/Dako Omnis was compared to (a) anti-PR clone PgR 636/Autostainer Link 48 (289 specimens) and to (b) anti-PR clone 16 (Leica Biosystems, Newcastle, UK) (144 specimens). In addition, the specificity of the ER and PR antibodies for Dako Omnis was evaluated on a set of normal tissue specimens.
Methods: Formalin-fixed, paraffin-embedded (FFPE) human breast carcinoma specimens and normal tissues were obtained from commercial providers or local hospitals. The specimens had no associated personal information and were not traceable back to the tissue donors. Tissue pretreatment and immunohistochemical staining were performed using the recommended protocol for each antibody and staining platform. The stained slides were evaluated for nuclear ER or PR expression according to ASCO/CAP guidelines (≥1% cut-off for positive) by pathologists who were blinded from the staining method and specimen ID. The concordance studies included breast cancer specimens covering the clinical range of ER or PR expression with approximately half the specimens in the negative (&lt;1%) category, and at least 10% of the specimens in the weakly positive (≥1 ≤10%) category in each study. Two-sided Wilson Score 95% Confidence Intervals were calculated using JMP software (SAS Institute, USA). For the analytical specificity studies the presence or absence of specific staining in the various normal tissue types was recorded.
Results: High concordance rates were observed with both anti-ERα clone EP1/Dako Omnis and anti-PR clone PgR 1294/Dako Omnis compared to the other ER/PR antibodies, with overall agreement rates exceeding 95% in all of the comparative studies. On a set of normal tissues, specific positive nuclear staining was observed only in tissue types known to express ERα or PR.
Conclusions: Monoclonal antibodies anti-ERα clone EP1 and anti-PR clone PgR 1294 configured as FLEX ready-to-use on Dako Omnis are sensitive and specific assays for detecting estrogen receptor and progesterone receptor in FFPE tissues. In comparison testing for assessment of hormonal receptor status on breast carcinoma specimens, anti-ERα clone EP1/Dako Omnis and anti-PR clone PgR 1294/Dako Omnis were highly concordant with commercially-available ER or PR antibodies.
Citation Format: Viale G, Dell'Orto P, Falzon M, Fält A, Hicks D, Hoff K, Jakobsen K, Jensen LB, Levy YY, McMahon L, Miller K, Russo L. Performance evaluation of two ready-to-use antibodies under development for the Dako Omnis automated staining platform on breast carcinoma specimens: Anti-estrogen receptor α clone EP1 and anti-progesterone receptor clone PgR 1294. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P1-01-16.