The human protein atlas blog

Proposed guidelines for antibody validation

Antibody Validation Mass Spectrometry

Graphical representation of the five proposed strategies

An international working group has formulated directives to evaluate antibodies used in biomedical research. A paper in Nature Methods suggests five conceptual pillars for antibody validation.

Antibodies are used frequently both in research and in the clinic, and today over two million commercial antibodies are available for the analysis of human proteins. This has been accompanied with a dramatic development in clinical treatment of diseases, like cancer and autoimmune diseases, where biological drugs in the form of antibodies have changed the treatment of many patient categories. At present, clinically used antibodies are under strict control, but antibodies used in research lack general accepted scientific guidelines for quality control. Lately, several papers have been published in international scientific journals, stating the questionable quality of many commercially available antibodies and they point out the problems this gives to life science research. .

In this context, an International Working Group on Antibody Validation (IWGAV), has been formed, consisting of researchers from institutions, such as Stanford University, Yale University, MIT, UCSD, University of Toronto, National Institutes of Health (NIH), European Molecular Biology Laboratory (EMBL), Niigata University in Japan and Science for Life Laboratory in Sweden. The aim is to establish standards for evaluation and quality control of antibodies, for both users and producers of antibodies.

– We recommend five basic strategies for validation of antibodies, says Mathias UhlÚn, professor at KTH Royal Institute of Technology and chair of IWGAV. We hope that these strategies will be used in the future both by researchers and by the antibody provider industry.

The five strategies outlined below are partly based on techniques developed recently, such as the DNA-editing method CRISPR-Cas9, and the aim is to provide firm evidence whether an antibody binds to its target protein in a specific manner.

  • Genetic strategies. The protein is knocked out using genetic methods.
  • Orthogonal strategies. An antibody-independent method is used.
  • Independent antibody strategies. Several antibodies are used.
  • Expression of tagged proteins. The target protein is tagged and analysed.
  • Mass spectrometry. The target protein is captured by an antibody and analysed.
  • These strategies for validation of antibodies will be discussed in the end of September at a meeting in California with representatives from the scientific community, international scientific journals and large public funders of biomedical research.

    Read the whole article on antibody validation in Nature Methods.

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