In this introduction to the topic, Professor Andreas Plückthun, University of Zürich, discusses the biology of the 3 principal types of commercial antibodies: polyclonal, monoclonal, and recombinant.  Surprisingly, only recombinants offer a reasonable hope of reproducibility in the long term – yet they remain very much the minority.

Professor Glenn Begley, Biocurate Pty Ltd, and Professor Cecilia Williams, KTH  Royal Institute of Technology, describe arduous, alarming and very costly experiences in failing to confirm “well established” critical data in oncology –a failure often driven by ineptly characterized antibodies.

Dr. Jan Voskuil, Aeonian Biotech, and Professor Andy Chalmers, University of Bath and CiteAb, delve into the mysterious world of original equipment manufacturers (OEMs), which can veil the identity of antibodies, and data-base repositories that help unveil which antibody has produced reliable results.

Professor Anita Bandrowski, University California San Diego and SciCrunch, and Dr. Jan Voskuil, Aeonian Biotech, probe how antibody reagents are identified.  If we can’t identify which antibody has been used, we can never reproduce experiments.  The Research Reagent Identifier systematically tags antibodies so they can be tracked, but the small-print on the data-sheets still hides many things.

Dr. Giovanna Roncador, Centro Nacional de Investigaciones Oncológicas and EuroMabNet, examinesthe criticality ofusing appropriate fit-for-purpose controls, both positive and negative, to validate antibodies in every experimental context.

Professors Aldrin Gomes and James Trimmer, both from University of California, Davis, dissect two “basic” validation technologies, Western blot and immunohistochemistry, to expose the very many things that can distort validation data.

Dr. Travis Hardcastle, Horizon, and Dr. Alejandra Solache, Abcam, describe the character of different gene-knockout technologies, one of the strongest validation technologies, and their value in large-scale antibody screening. 

Dr. Mike Taussig, Cambridge Protein Arrays, and Dr. Fridtjof Lund-Johansen, Oslo University Hospital, look at array and immunoprecipitation-mass spectrometry technologies, which offer a broader and deeper image of antibody specificity and selectivity for validation, than classical validation technologies.

Dr. Andrew Bradbury, Specifica, suggests ways out of the validation maze by rigorous molecular identification of recombinant tool antibodies. This achievable goal could eliminate many of the shadowy issues described in this series of webinars.

Future recombinant binding tools may avoid antibodies, and use diversified libraries based on small and versatile protein scaffolds, for example the DARPins (Designed Ankyrin Repeat Proteins).  DARPins are single-chain, monodisperse, and non-immunogenic, and can be made rapidly and cheaply in bacteria in many formats.  They can also be expressed in and on cells as non-aggregating fusion proteins with tunable valencies, which opens a wide range of previously inaccessible approaches in cell biology and biochemistry.