Nat. Meth. doi:10.1038/nmeth.4632 (2018)

There is a well-documented reproducibility crisis in the biomedical sciences, and an important contributor to this is the widespread use of poorly validated and unstandardized monoclonal antibodies (mAbs). In Nature Methods, Blackshaw and colleagues describe a robust screening and validation pipeline that allows the generation of high-quality mouse mAbs to 737 human transcription factors. Full-length antigens or their domains are used to generate mAbs that are then initially screened on a human protein array. mAbs that pass the initial screen are then subjected to secondary validation via various applications, including immunoprecipitation, immunoblot analysis and, for a subset, immunohistochemistry and chromatin immunoprecipitation followed by deep sequencing. A meta-analysis of the pipeline helps to identify factors that influence the probability of generating high-quality mAbs, such as the route of immunization.