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A New Rabbit Monoclonal Antibody (4B5) for the Immuno-histochemical (IHC) Determination of the HER2 Status in Breast Cancer: Comparison With CB11, Fluorescence In Situ Hybridization (FISH), and Interlaboratory Reproducibility

A New Rabbit Monoclonal Antibody (4B5) for the Immuno-histochemical (IHC) Determination of the... LETTER TO THE EDITOR A New Rabbit TABLE 1. Recommended Staining Protocols for PATHWAY anti-HER-2/neu (4B5) With iVIEW DAB Detection Kit Monoclonal Antibody Platform or Method (4B5) for the Immuno- Procedure Type NexES IHC BenchMark Series Deparaffinization Off Line (manual) Selected histochemical (IHC) Cell Conditioning Off Line (manual), Cell Conditioning 1, 2 min, Cell Conditioning 1, (Antigen Unmasking) Decloaking Chamber, 1201C Standard Determination of the Enzyme (Protease) None required None required Antibody (Primary) Approximately 32 min, 371C Approximately HER2 Status in Breast 32 min, 371C A/B Block (Biotin Required Required Cancer: Comparison Blocking) Counterstain Hematoxylin II, 4 min Hematoxylin II, With CB11, (Hematoxylin) 4 min Postcounterstain Bluing, 4 min Bluing, 4 min Fluorescence In Situ IHC indicates immunohistochemistry. Hybridization (FISH), and Interlaboratory antibody is Ventana’s basic pH Cell vided in the package insert. This Reproducibility Conditioning 1 solution (CC1, Catalog limitation of use is specifically called no. 950–124). This antigen unmasking out in the package insert under solution contains a Tris-based buffer ‘‘LIMITATIONS (General limitation To the Editor: (in contrast to the solution described number 5).’’ Specifically, the package We welcome the comments made by Egervari et al; see below), and use of insert states ‘‘Ventana provides anti- by Kristof Egervari, Zoltan Szollosi, CC1 is clearly called out in the package bodies and reagents at optimal dilu- MD, PhD, and Zoltan Nemes, MD, insert as part of the recommended tion for use when the provided DSci from the Department of Pathol- staining protocol for the PATHWAY instructions are followed. Any devia- ogy at the Medical and Health Science anti-HER-2/neu (4B5)RabbitMono- tion from recommended test proce- Center, University of Debrecen, Hun- clonal Primary Antibody. dures may invalidate expected results. gary regarding our paper ‘‘A New Egervari et al state that ‘‘[t]o Appropriate controls must be em- Rabbit Monoclonal Antibody (4B5) unmask the epitopes [they] used the ployed and documented. Users who for the Immunohistochemical (IHC) conventional high pressure antigen re- deviate from recommended test pro- Determination of the HER2 Status trieval method in citrate buffer (pH 6.5, cedures must accept responsibility for in Breast Cancer: Comparison with for 3 min), as recommended by the interpretation of patient results.’’ CB11, fluorescence in situ hybridiza- supplier.’’ As alluded to above, citrate We strongly encourage Egervari tion (FISH), and interlaboratory re- buffer is not the recommended retrieval et al and all users of the PATHWAY producibility,’’ recently published in solution described in the Instructions anti-HER-2/neu (4B5) Rabbit Mono- Applied Immunohistochemistry and Mo- for Use in the package insert for clonal Primary Antibody to read and lecular Morphology (Appl Immunohis- PATHWAY anti-HER-2/neu (4B5) to adhere to the instructions provided tochem Mol Morphol. 2007;15:94–102) Rabbit Monoclonal Primary Antibody. in the package insert. This will ensure and appreciate the opportunity to Table 1 above is from the package that standardized and appropriate respond. insert, and clearly identifies CC1 as the staining results are obtained with this One goal at Ventana Medical recommended reagent for antigen un- antibody. Systems is to increase the reproduci- masking and 2 min as the recommended bility and reliability of immunohisto- time for cell conditioning using a high William C. Powell, PhD* chemical staining through the highest pressure Decloaking Chamber before Patrick C. Roche, PhD* quality reagents with the latest in staining on a NexES instrument. Raymond R. Tubbs, DOww automation. As part of the develop- The anti-HER-2/neu (4B5) Rab- *Reagent Development ment process for our PATHWAY bit Monoclonal Primary Antibody Ventana Medical Systems Inc anti-HER-2/neu (4B5) Rabbit Mono- product has received premarket Tucson, AZ clonal Primary Antibody, we deter- approval from the Food and Drug wDepartment of Molecular Pathology mined that the optimal antigen Administration and is to be used in Cleveland Clinic unmasking solution for use with this accordance with the instructions pro- Cleveland, OH Appl Immunohistochem Mol Morphol Volume 16, Number 6, December 2008 569 http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Applied Immunohistochemistry & Molecular Morphology Wolters Kluwer Health

A New Rabbit Monoclonal Antibody (4B5) for the Immuno-histochemical (IHC) Determination of the HER2 Status in Breast Cancer: Comparison With CB11, Fluorescence In Situ Hybridization (FISH), and Interlaboratory Reproducibility

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/lp/wolters-kluwer-health/a-new-rabbit-monoclonal-antibody-4b5-for-the-immuno-histochemical-ihc-1snjCXi1i0
ISSN
1541-2016
DOI
10.1097/PAI.0b013e3181895d6c
pmid
18931613
Publisher site
See Article on Publisher Site

Abstract

LETTER TO THE EDITOR A New Rabbit TABLE 1. Recommended Staining Protocols for PATHWAY anti-HER-2/neu (4B5) With iVIEW DAB Detection Kit Monoclonal Antibody Platform or Method (4B5) for the Immuno- Procedure Type NexES IHC BenchMark Series Deparaffinization Off Line (manual) Selected histochemical (IHC) Cell Conditioning Off Line (manual), Cell Conditioning 1, 2 min, Cell Conditioning 1, (Antigen Unmasking) Decloaking Chamber, 1201C Standard Determination of the Enzyme (Protease) None required None required Antibody (Primary) Approximately 32 min, 371C Approximately HER2 Status in Breast 32 min, 371C A/B Block (Biotin Required Required Cancer: Comparison Blocking) Counterstain Hematoxylin II, 4 min Hematoxylin II, With CB11, (Hematoxylin) 4 min Postcounterstain Bluing, 4 min Bluing, 4 min Fluorescence In Situ IHC indicates immunohistochemistry. Hybridization (FISH), and Interlaboratory antibody is Ventana’s basic pH Cell vided in the package insert. This Reproducibility Conditioning 1 solution (CC1, Catalog limitation of use is specifically called no. 950–124). This antigen unmasking out in the package insert under solution contains a Tris-based buffer ‘‘LIMITATIONS (General limitation To the Editor: (in contrast to the solution described number 5).’’ Specifically, the package We welcome the comments made by Egervari et al; see below), and use of insert states ‘‘Ventana provides anti- by Kristof Egervari, Zoltan Szollosi, CC1 is clearly called out in the package bodies and reagents at optimal dilu- MD, PhD, and Zoltan Nemes, MD, insert as part of the recommended tion for use when the provided DSci from the Department of Pathol- staining protocol for the PATHWAY instructions are followed. Any devia- ogy at the Medical and Health Science anti-HER-2/neu (4B5)RabbitMono- tion from recommended test proce- Center, University of Debrecen, Hun- clonal Primary Antibody. dures may invalidate expected results. gary regarding our paper ‘‘A New Egervari et al state that ‘‘[t]o Appropriate controls must be em- Rabbit Monoclonal Antibody (4B5) unmask the epitopes [they] used the ployed and documented. Users who for the Immunohistochemical (IHC) conventional high pressure antigen re- deviate from recommended test pro- Determination of the HER2 Status trieval method in citrate buffer (pH 6.5, cedures must accept responsibility for in Breast Cancer: Comparison with for 3 min), as recommended by the interpretation of patient results.’’ CB11, fluorescence in situ hybridiza- supplier.’’ As alluded to above, citrate We strongly encourage Egervari tion (FISH), and interlaboratory re- buffer is not the recommended retrieval et al and all users of the PATHWAY producibility,’’ recently published in solution described in the Instructions anti-HER-2/neu (4B5) Rabbit Mono- Applied Immunohistochemistry and Mo- for Use in the package insert for clonal Primary Antibody to read and lecular Morphology (Appl Immunohis- PATHWAY anti-HER-2/neu (4B5) to adhere to the instructions provided tochem Mol Morphol. 2007;15:94–102) Rabbit Monoclonal Primary Antibody. in the package insert. This will ensure and appreciate the opportunity to Table 1 above is from the package that standardized and appropriate respond. insert, and clearly identifies CC1 as the staining results are obtained with this One goal at Ventana Medical recommended reagent for antigen un- antibody. Systems is to increase the reproduci- masking and 2 min as the recommended bility and reliability of immunohisto- time for cell conditioning using a high William C. Powell, PhD* chemical staining through the highest pressure Decloaking Chamber before Patrick C. Roche, PhD* quality reagents with the latest in staining on a NexES instrument. Raymond R. Tubbs, DOww automation. As part of the develop- The anti-HER-2/neu (4B5) Rab- *Reagent Development ment process for our PATHWAY bit Monoclonal Primary Antibody Ventana Medical Systems Inc anti-HER-2/neu (4B5) Rabbit Mono- product has received premarket Tucson, AZ clonal Primary Antibody, we deter- approval from the Food and Drug wDepartment of Molecular Pathology mined that the optimal antigen Administration and is to be used in Cleveland Clinic unmasking solution for use with this accordance with the instructions pro- Cleveland, OH Appl Immunohistochem Mol Morphol Volume 16, Number 6, December 2008 569

Journal

Applied Immunohistochemistry & Molecular MorphologyWolters Kluwer Health

Published: Dec 1, 2008

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