introduction to the ventana p40 (bc28) mouse monoclonal primary antibody: focus on performance and...

p40 (BC28) Mouse Monoclonal Antibody

Comprehensive Overview – May 2015

Introduction to lung cancer

Diagnostic Algorithm

p40 background and product details

p40 Publication Support

p40 (BC28) Basic Information

Lung cancer facts

1.8 million of all cancers

new cases

12.7% 1st Mortality:

3rd Incidence:

World statistics

5-year survival of NSCLC is as low as 16%

Source: globocan.iarc.fr/old/FactSheets/cancers/lung-new.asp

15%

5%

15%

40%

25%

85%

Introduction to lung cancer Lung Cancer Subtypes

Shaffer, Anita T. Expanding TKI Therapy in Advanced Lung Cancer. OncLive. Published Online: Wednesday, October 9, 2013. http://www.onclive.com/publications/oncology-live/2013/september-2013/expanding-tki-therapy-in-advanced-lung-cancer/2 Medicine.net. Lung Cancer. Accessed 11DEC2014. http://www.medicinenet.com/lung_cancer/page7.htm.

Stratifying lung cancer Critical focus: Non-Small Cell Lung Carcinoma (NSCLC)

2

1.  Rossi, et al. Subtyping Non-Small Cell Lung Cancer. INT J SURG PATHOL. 21:326.2013. 2.  Tacha D, Yu C, Bremer R, Qi W, Haas T. A 6-antibody panel for the classification of lung adenocarcinoma versus squamous cell carcinoma. Appl Immunohistochem Mol Morphol. 2012;20:201-207.

+

TTF-1 + Napsin A +

p40 + CK 5/6 +

Subtyping Non-Small Cell Lung Cancer Adenocarcinoma vs. squamous cell carcinoma

v  It is critical for downstream clinical decisions to first accurately differentiate between squamous cell carcinomas and adenocarcinomas

v  None of these antibodies are 100% specific or 100% sensitive alone

v  p63 (4A4) has been replaced by p40 (BC28) in this panel

v  This 4 antibody panel is supported by peer reviewed literature* and may help ensure quick and accurate subtyping of NSCLC

*Wei Zhao, Hui Wang, Yan Peng, Bo Tian, Lei Peng, Da-Chuan Zhang. ∆Np63, CK5/6, TTF-1 and napsin A, a reliable panel to subtype non-small cell lung cancer in biopsy specimens. Int J Clin Exp Pathol 2014;7(7):4247-4253

Thyroid Transcription Factor-1 (SP141) Rabbit Monoclonal Primary Antibody

Cytokeratin 5/6 (D5/16B4) Mouse Monoclonal Primary Antibody

Napsin A (MRQ-60) Mouse Monoclonal Primary Antibody

p40 (BC28) Mouse Monoclonal Primary Antibody

The Product

Class I/CE-IVD mouse monoclonal antibody targeting the p40 epitope utilizing the BC28 clone developed with OptiView DAB IHC Detection and ultraView Detection in a ready-to-use dispenser on BenchMark GX, XT, and ULTRA IHC/ISH instruments, for primary use in lung cancer

The Science

•  p63 is a ‘two-in-one’ family of opposite molecules: TA– a p53-like tumor suppressor and ∆Np63–an oncogene. p40 antibody recognizes an epitope which is unique to ∆Np63

•  BC28 targets amino acids 5-17 of the p40 epitope and demonstrates nuclear staining •  Utility: squamous differentiation in lung, bladder, breast, prostate and head & neck cancers2

The Value

•  p40 is roughly equivalent to p63 in sensitivity for squamous cell carcinoma, but it is markedly superior to p63 in specificity (98% vs. 60%)1

•  Routine use of p40 in place of p63 for the differential diagnosis of adenocarcinoma vs. squamous cell carcinoma is well supported1-4

p40 (BC28) Mouse Monoclonal Primary Antibody Summary of key product details

1.  Bishop, et al. p40 (∆Np63) is superior to p63 for the diagnosis of pulmonary squamous cell carcinoma. Modern Pathology. 2012. 2.  Tacha, et al. An Immunohistochemical Analysis of a Newly Developed, Mouse Monoclonal p40 (BC28) Antibody in Lung, Bladder, Skin, Breast, Prostate, and Head and Neck Cancers. Arch Pathol Lab Med—Vol 138, October 2014. 3.  Nonaka, Daisuke MD. Study of ΔNp63 Expression in Lung Non-Small Cell Carcinomas. Am J Surg Pathol 2012; 36:895–899. 4.  Tatsumori et al. p40 is the Best Marker for Diagnosing Pulmonary Squamous Cell Carcinoma. Appl Immunohistochem Mol Morphol 2014; 22:377–382.

Staining results by IHC/ISH platform Lung Squamous Cell Carcinoma stained with p40 (BC28) assay

BenchMark GX BenchMark XT

BenchMark ULTRA

Examples stained with OptiView DAB IHC Detection

Skin Breast Prostate

Lung SCC Breast ductal carcinoma Undifferentiated carcinoma

Tissue Staining Examples Stained with p40 (BC28) antibody with OptiView DAB IHC Detection

p40 (BC28) vs. p63 (4A4) staining results Lung Squamous Cell Carcinoma and Adenocarcinoma

Squamous cell carcinoma stained positive with p40 (BC28) assay using OptiView DAB IHC detection

Squamous cell carcinoma stained positive with p63 (4A4) assay using ultraView DAB IHC detection

Adenocarcinoma stained negative with p40 (BC28) assay using OptiView DAB IHC detection

Adenocarcinoma stained positive with the p63 (4A4) assay using ultraView DAB IHC detection

The use of p40 is recommended as a replacement for p63 when subclassifying non-small cell lung cancer to avoid falsely classifying p63+ adenocarcinoma as a squamous cell carcinoma.*

Bishop, Justin A, Julie Teruya-Feldstein, William H Westra, Giuseppe Pelosi, William D Travis and Natasha Rekhtman. p40 (∆Np63) is superior to p63 for the diagnosis of pulmonary squamous cell carcinoma. Modern Pathology. 2012.

1.  Zhao, et al. ∆Np63, CK5/6, TTF-1 and napsin A, a reliable panel to subtype non-small cell lung cancer in biopsy specimens. Int J Clin Exp Pathol 2014;7(7):4247-4253

2.  Bishop, et al. p40 (∆Np63) is superior to p63 for the diagnosis of pulmonary squamous cell carcinoma. Modern Pathology. 2012. 3.  Butnor, et al. [1876] p40 (ΔNp63) and Keratin 34βE12 Provide Greater Diagnostic Accuracy Than p63 in the Evaluation of Small Cell Lung Carcinoma in Small

Biopsy Samples. USCAP Abstract. 2013. 4.  Chen, et al. [351] The Role of p40 Immunostain in the Cytological Differential Diagnosis of Squamous Cell Carcinoma and Adenocarcinoma of the Lung. USCAP

Abstract. 2013. 5.  Gailey, et al. [410] Differentiating Small Cell Carcinoma from Squamous Cell Carcinoma on Fine Needle Aspiration: A Head-To-Head Comparison of p40 and

p63. USCAP Abstract. 2014. 6.  Liao, et al. Sensitivity and Specificity of p40, p63, and SOX2 Monoclonal Antibodies in Lung Squamous Cell Carcinoma.Poster. 2013. 7.  Tacha, et al. An Immunohistochemical Analysis of a Newly Developed, Mouse Monoclonal p40 (BC28) Antibody in Lung, Bladder, Skin, Breast, Prostate, and

Head & Neck Cancers. Arch Pathol Lab. Oct 2014. 8.  Nonaka, Daisuke MD. Study of ΔNp63 Expression in Lung Non-Small Cell Carcinomas. Am J Surg Pathol 2012; 36:895–899. 9.  Pelosi et al. Δp63 (p40) and Thyroid Transcription Factor-1 Immunoreactivity on Small Biopsies or Cellblocks for Typing Non-small Cell Lung Cancer A Novel

Two-Hit, Sparing-Material Approach. Jo Thor Feb 2012. 10.  Tatsumori et al. p40 is the Best Marker for Diagnosing Pulmonary Squamous Cell Carcinoma: Comparison with p63, Cytokeratin 5/6, Desmocollin-3, and Sox2.

Appl Immunohistochem Mol Morphol 2014; 22:377–382. 11.  Vogt, et al. [436] p40 (ΔNp63): A Highly Sensitive and Specific Immunohistochemical (IHC) Marker for Diagnosing Pulmonary Squamous Cell Carcinomas

(SQCC) in Fine Needle Aspirates. USCAP Abstract. 2013. 12.  Sailer, Verena et al. Comparison of p40 (DNp63) and p63 expression in prostate tissues – which one is the superior diagnostic marker for basal cells?

Histopathology. 2013. 63: 50–56. 13.  Bishop, et al. Use of p40 and p63 Immunohistochemistry and Human Papillomavirus Testing as Ancillary Tools for the Recognition of Head and Neck

Sarcomatoid Carcinoma and Its Distinction From Benign and Malignant Mesenchymal Processes. Am J Surg Pathol. 2014. 38:257–264. 14.  Rooper, Lisa, Rajni Sharma, Justin A. Bishop. Polymorphous Low Grade Adenocarcinoma has a Consistent p63+/p40- Immunophenotype that Helps Distinguish

it from Adenoid Cystic Carcinoma and Cellular Pleomorphic Adenoma. Head and Neck Pathol. 2014. DOI 10.1007/s12105-014-0554-4 15.  Brustmann, Hermann. p40 as a Basal Cell Marker in the Diagnosis of Prostate Glandular Proliferations: A Comparative Immunohistochemical Study with

34betaE12. Hindawi Publishing Corporation Pathology Research International. Volume 2015, Article ID 897927, 5 pages 16.  Righi, et al. Impact of Non–Small-Cell Lung Cancer-Not Otherwise Specified Immunophenotyping on Treatment Outcome. J Thorac Oncol. 2014;9: 1540–1546.

Key p40 support publications Over 40 publications since 2012

p40 Publication Summary Example 1 Publication: Tacha, David PhD; Ryan Bremer, PhD; Thomas Haas, DO; Weiman Qi, PhD, MD. An Immunohistochemical Analysis of a Newly Developed, Mouse Monoclonal p40 (BC28) Antibody in Lung, Bladder, Skin, Breast, Prostate and Head and Neck Cancers. Arch Pathol Lab Med—Vol 138, October 2014. The major goal of this study was to evaluate the specificity and sensitivity of p40 (BC28) mouse monoclonal primary antibody in normal and neoplastic tissues with emphasis on lung cancer. The p40 (BC28) antibody was also evaluated in breast, bladder, skin, prostate, and head and neck cancers. The staining characteristics of p40 (BC28), p63 (4A4) and p40 (polyclonal) antibodies were compared in different types of lung cancers. In addition, p40 (BC28) antibody was evaluated in benign prostate glands and in prostatic intraepithelial neoplasia and compared with the p63 (4A4) antibody. The results demonstrate the superiority of p40 (BC28) compared to p63 (4A4) in the differentiation of lung tumors. The data indicate that p40 (BC28) has sensitivity equal to that of p63 (4A4) in lung squamous cell carcinoma, however fewer lung adenocarcinoma cases were p40 positive, supporting a claim of increased specificity of p40 (BC28) over p63 (4A4). Furthermore, notably superior staining was observed with p40 (BC28) compared to p40 (polyclonal) in lung cancer cases, where p40 (polyclonal) displayed nonspecific, cytoplasmic staining. In addition, the data in this study provide the evidence for the use of p40 (BC28) as a high-quality antibody for determining squamous cell carcinoma in lung, skin, head and neck and also in urothelial carcinomas. p40 (BC28) displayed cleaner and more intense staining pattern when compared to p63 in basal cells in prostate tissue.

Items herein are the opinion of Ventana Medical Systems, Inc.

p40 Publication Summary Example 2 Publication: Wei Zhao, Hui Wang, Yan Peng, Bo Tian, Lei Peng, Da-Chuan Zhang. ∆Np63, CK5/6, TTF-1 and napsin A, a reliable panel to subtype non-small cell lung cancer in biopsy specimens. Int J Clin Exp Pathol 2014; 7(7):4247-4253. The aim of this study was to evaluate a panel of different IHC markers in subtyping poorly differentiated NSCLC. There were 48 cases included in the study that could not be classified by H&E alone. The markers evaluated in this study included p40 (ΔNp63), CK 5/6 (D5/16B4), TTF-1 (8G7G3/1), Napsin A (MRQ-60), p63 (4A4), CK7 (OV-TL12/30), CK8/18 (5D3) and keratin 34βE12. The study showed that a panel including p40 (polyclonal), CK5/6 (D5/16B4), TTF-1 (8G7G3/1), and Napsin A (MRQ-60) correctly classified 39 of 48 cases (81%) of poorly differentiated NSCLC. Nine tumors could not be further classified. The inclusion of p40 (polyclonal) as a substitute for p63 (4A4) in this panel was based on findings that p40 provided a higher specificity than p63 for lung adenocarcinoma and equal sensitivity for lung squamous cell carcinoma. CK7, CK8/18 and 34βE12 were excluded from the panel due to low specificity of CK7 and CK8/18 for adenocarcinoma and 34βE12 for SCC. In conclusion, the proposed IHC panel for lung cancer classification includes the following markers - p40, CK5/6, TTF1, and Napsin A.

Items herein are the opinion of Ventana Medical Systems, Inc.

p40 Publication Summary Example 3 Publication: Bishop, Justin A, Julie Teruya-Feldstein, William H Westra, Giuseppe Pelosi, William D Travis and Natasha Rekhtman. p40 (∆Np63) is superior to p63 for the diagnosis of pulmonary squamous cell carcinoma. Modern Pathology. 2012. The purpose of this study was to compare the specificity and sensitivity of the p40 (polyclonal) antibody to that of a p63 (4A4) antibody in different lung tumors and large cell lymphoma. Overall, 470 cases including 81 lung squamous cell carcinomas, 237 lung adenocarcinomas, and 152 large cell lymphomas were evaluated in this study. The data indicate that although the sensitivity of p40 in squamous cell carcinoma is comparable to p63 (4A4) antibody, the specificity of p40 is significantly better than p63 (4A4). These results provide support for the routine use of p40 instead of p63 for the diagnosis of pulmonary squamous cell carcinoma. Additional publication summaries are available on the: p40 (BC28) Mouse Monoclonal Primary Antibody Abstract summary sheet Items herein are the opinion of Ventana Medical Systems, Inc.

Catalogue number 790-4950 / 07394420001

Type Mouse Monoclonal – IgG1

Clone BC28

Number of tests 50 tests

Reactivity Paraffin (10% NBF-fixed recommended)

Localization Nuclear

Control sample Lung squamous cell carcinoma

Regulatory status CE-IVD, Class I

Recommended protocol

BenchMark GX* BenchMark XT BenchMark ULTRA

OptiView •  CC1, 32 min •  16 min Ab inc @ 37°C •  Hematoxylin II, 4 minute •  Bluing, 4 minutes

•  CC1, 32 min •  16 min Ab inc @ 37°C •  Hematoxylin II, 4 minute •  Bluing, 4 minutes

•  CC1, 32 min •  16 min Ab inc @ 36°C •  Hematoxylin II, 4 minute •  Bluing, 4 minutes

ultraView •  CC1, standard •  16 min Ab inc @ 37°C •  Amplification •  Hematoxylin II, 4 minute •  Bluing, 4 minutes

•  CC1, standard •  16 min Ab inc @ 37°C •  Amplification •  Hematoxylin II, 4 minute •  Bluing, 4 minutes

•  CC1, standard •  16 min Ab inc @ 36°C •  Amplification •  Hematoxylin II, 4 minute •  Bluing, 4 minutes * BenchMark GX Platform not available in the US

p40 (BC28) Mouse Monoclonal Primary Antibody

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VENTANA, the VENTANA logo, BENCHMARK, OPTIVIEW and ultraView are trademarks of Roche. All other trademarks are the property of their respective owners. 5627A-11 0415

introduction to the ventana p40 (bc28) mouse monoclonal primary antibody: focus on performance and...

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