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Validation of specificity of antibodies for immunohistochemistry: the case of ROR2

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An Erratum to this article was published on 29 October 2016

Abstract

The Wnt signalling receptor receptor tyrosine kinase-like orphan receptor 2 (ROR2) is implicated in numerous human cancers. However, there have been conflicting reports regarding ROR2 expression, some studies showing upregulation and others downregulation of ROR2 in the same cancer type. The majority of these studies used immunohistochemistry (IHC) to detect ROR2 protein, without validation of the used antibodies. There appears to be currently no consensus on the antibody best suited for ROR2 detection or how ROR2 expression changes in various cancer types. We examined three commercially available ROR2 antibodies and found that only one bound specifically to ROR2. Another antibody cross-reacted with other proteins, and the third failed to detect ROR2 at all. ROR2 detection by IHC on 107 patient samples using the ROR2 specific antibody showed that the majority of colorectal cancers show loss of ROR2 protein. We found no association between ROR2 staining and poor patient survival, as had been previously reported. These results question the previously reported association between ROR2 and poor patient survival in colorectal cancer. Future studies should use fully validated antibodies when detecting ROR2 protein, as non-specific staining can lead to irrelevant observations and misinterpretations.

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Authors and Affiliations

  1. Adult Cancer Program, Level 2, Metastasis Research Group, Lowy Cancer Research Centre and School of Women’s and Children’s Health, University of New South Wales, Sydney, NSW, 2052, Australia

    Sean S.Q. Ma, Claire E. Henry, Estelle Llamosas, Rupert Higgins & Caroline E. Ford

  2. Faculty of Medicine, Medicines Policy Research Unit, Centre for Big Data Research in Health, UNSW, Sydney, Australia

    Benjamin Daniels

  3. Colorectal Cancer Group, Adult Cancer Program, Lowy Cancer Research Centre, UNSW, Sydney, Australia

    Luke B. Hesson

  4. University of Queensland, St. Lucia, Australia

    Nicholas J. Hawkins & Robyn L. Ward

Authors
  1. Sean S.Q. Ma
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  2. Claire E. Henry
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  3. Estelle Llamosas
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  4. Rupert Higgins
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  5. Benjamin Daniels
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  6. Luke B. Hesson
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  7. Nicholas J. Hawkins
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  8. Robyn L. Ward
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  9. Caroline E. Ford
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Corresponding author

Correspondence to Caroline E. Ford.

Ethics declarations

Ethics approval was obtained for all research performed on patient samples. The ethics committee approval numbers are HREC 00113, H00/022 and H04/024, UNSW ethics committee. Informed consent was obtained from all patients participating in the study.

Funding

This work was supported by the NSW Cancer Council. Support and funding was also provided by the Translational Cancer Research Network (TCRN) and a UNSW Medicine Faculty Research Grant.

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

An erratum to this article is available at http://dx.doi.org/10.1007/s00428-016-2036-4.

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Ma, S.S., Henry, C.E., Llamosas, E. et al. Validation of specificity of antibodies for immunohistochemistry: the case of ROR2. Virchows Arch 470, 99–108 (2017). https://doi.org/10.1007/s00428-016-2019-5

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  • DOI: https://doi.org/10.1007/s00428-016-2019-5

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