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Virchows Archiv

, Volume 462, Issue 3, pp 269–279 | Cite as

Validation of the BRCA1 antibody MS110 and the utility of BRCA1 as a patient selection biomarker in immunohistochemical analysis of breast and ovarian tumours

  • Roy Milner
  • Helen Wombwell
  • Sonia Eckersley
  • Donna Barnes
  • Juli Warwicker
  • Erica Van Dorp
  • Simon Dearden
  • Glen Hughes
  • Chris Harbron
  • Bob Wellings
  • Darren Hodgson
  • Chris Womack
  • Neil Gray
  • Alan Lau
  • Mark J. O’Connor
  • Catherine Marsden
  • Alexander J. KvistEmail author
Original Article

Abstract

BRCA1 protein measurement has previously been evaluated as a potential diagnostic marker without reaching a conclusive recommendation. In this study, we applied current best practice in antibody validation to further characterize MS110, a widely used antibody targeting BRCA1. Antibody specificity was investigated using different biochemical validation techniques. We found that BRCA1 could not be reliably detected using immunoprecipitation and Western blot in endogenously expressing cells. We used immunohistochemistry on formalin-fixed paraffin-embedded cell pellets to establish compatibility with formalin-fixed paraffin-embedded samples. We demonstrated that in transfected cells and cell lines with known genetic BRCA1 status, MS110 successfully detected BRCA1 giving the expected level of staining in immunohistochemistry. Following this, we investigated the use of BRCA1 protein measurement by immunohistochemistry in a cohort of triple negative breast and serous ovarian tumour samples to explore the use of BRCA1 protein measurement by immunohistochemistry for patient stratification. Using MS110 in repeated standardized experiments, on serial sections from a panel of patient samples, results demonstrated considerable run-to-run variability. We concluded that in formalin-fixed tissue samples, MS110 does detect BRCA1; however, using standard methodologies, BRCA1 expression levels in tissue samples is incompatible with the use of this protein as a statistically robust patient selection marker in immunohistochemistry. These results demonstrate the need for further development to deliver BRCA1 protein quantification by immunohistochemistry as a patient stratification marker.

Keywords

MS110 BRCA1 Antibody validation Patient selection Breast cancer 

Notes

Acknowledgments

All research in the manuscript was funded by AstraZeneca PLC. We would like to thank Dr. Madhuri Warren of Pathology Diagnostics, Ltd. for the meticulous work of second pathologist scoring of all immunohistochemical samples.

Conflicts of interest

At the time of data collection and writing of the manuscript all authors, with the exception of Erica Van Dorp, were employees of AstraZeneca R&D. Erica Van Dorp declares she has no conflict of interest.

Supplementary material

428_2012_1368_Fig5_ESM.jpg (2.7 mb)
Supplementary Fig. 1

Immunohistochemical staining of normal breast tissue for BRCA1 and PARP1. Panel a shows staining for BRCA1 on normal breast tissue with MS110. Panel b shows staining for PARP on normal breast tissue with Serotec antibody MCA1522. Scale bar, 100 μm (JPEG 2717 kb)

428_2012_1368_MOESM1_ESM.eps (5.5 mb)
High resolution (EPS 5620 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Roy Milner
    • 1
  • Helen Wombwell
    • 1
  • Sonia Eckersley
    • 1
  • Donna Barnes
    • 1
  • Juli Warwicker
    • 1
  • Erica Van Dorp
    • 1
  • Simon Dearden
    • 1
  • Glen Hughes
    • 1
  • Chris Harbron
    • 1
  • Bob Wellings
    • 1
  • Darren Hodgson
    • 1
  • Chris Womack
    • 1
  • Neil Gray
    • 1
  • Alan Lau
    • 1
  • Mark J. O’Connor
    • 1
  • Catherine Marsden
    • 1
  • Alexander J. Kvist
    • 1
    • 2
    Email author
  1. 1.AstraZenecaMacclesfieldUK
  2. 2.MacclesfieldUK

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