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Insufficient antibody validation challenges oestrogen receptor beta research

Author

Listed:
  • Sandra Andersson

    (Genetics and Pathology, Uppsala University, Science for Life Laboratory)

  • Mårten Sundberg

    (Uppsala University, Science for Life Laboratory)

  • Nusa Pristovsek

    (Genetics and Pathology, Uppsala University, Science for Life Laboratory)

  • Ahmed Ibrahim

    (School of Biotechnology, Science for Life Laboratory, KTH Royal Institute of Technology
    National Research Centre)

  • Philip Jonsson

    (University of Houston
    Present address: Department of Epidemiology and Biostatistics and Human Oncology and Pathogenesis Program, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA)

  • Borbala Katona

    (Genetics and Pathology, Uppsala University, Science for Life Laboratory)

  • Carl-Magnus Clausson

    (Genetics and Pathology, Uppsala University, Science for Life Laboratory)

  • Agata Zieba

    (Genetics and Pathology, Uppsala University, Science for Life Laboratory)

  • Margareta Ramström

    (Uppsala University, Science for Life Laboratory)

  • Ola Söderberg

    (Uppsala University)

  • Cecilia Williams

    (School of Biotechnology, Science for Life Laboratory, KTH Royal Institute of Technology
    University of Houston
    Karolinska Institutet)

  • Anna Asplund

    (Genetics and Pathology, Uppsala University, Science for Life Laboratory)

Abstract

The discovery of oestrogen receptor β (ERβ/ESR2) was a landmark discovery. Its reported expression and homology with breast cancer pharmacological target ERα (ESR1) raised hopes for improved endocrine therapies. After 20 years of intense research, this has not materialized. We here perform a rigorous validation of 13 anti-ERβ antibodies, using well-characterized controls and a panel of validation methods. We conclude that only one antibody, the rarely used monoclonal PPZ0506, specifically targets ERβ in immunohistochemistry. Applying this antibody for protein expression profiling in 44 normal and 21 malignant human tissues, we detect ERβ protein in testis, ovary, lymphoid cells, granulosa cell tumours, and a subset of malignant melanoma and thyroid cancers. We do not find evidence of expression in normal or cancerous human breast. This expression pattern aligns well with RNA-seq data, but contradicts a multitude of studies. Our study highlights how inadequately validated antibodies can lead an exciting field astray.

Suggested Citation

  • Sandra Andersson & Mårten Sundberg & Nusa Pristovsek & Ahmed Ibrahim & Philip Jonsson & Borbala Katona & Carl-Magnus Clausson & Agata Zieba & Margareta Ramström & Ola Söderberg & Cecilia Williams & An, 2017. "Insufficient antibody validation challenges oestrogen receptor beta research," Nature Communications, Nature, vol. 8(1), pages 1-14, August.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_ncomms15840
    DOI: 10.1038/ncomms15840
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