Insufficient antibody validation challenges oestrogen receptor beta research

Sandra Andersson, Mårten Sundberg, Nusa Pristovsek, Ahmed Ibrahim, Philip Jonsson, Borbala Katona, Carl-Magnus Clausson, Agata Zieba, Margareta Ramström, Ola Söderberg, Cecilia Williams () and Anna Asplund
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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
Philip Jonsson: University of Houston
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
Anna Asplund: Genetics and Pathology, Uppsala University, Science for Life Laboratory

Nature Communications, 2017, vol. 8, issue 1, 1-14

Abstract: 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.

Date: 2017
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DOI: 10.1038/ncomms15840

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