[Letter to the Editor] The Need for Improved Education and Training in Research Antibody Usage and Validation Practices

Abstract

Address correspondence to Leonard P. Freedman, Global Biological Standards Institute, 1020 19th Street, NW, Suite 550, Washington, DC, 20036. E-mail: lfreedman@gbsi.org.

Reproducibility2020: Progress and priorities

“…The research community has acknowledged that antibodies are an area of widespread error and inaccuracy 23 . The Antibody Validation Initiative, involving stakeholders throughout the research community and led by GBSI, is an example that could be replicated in other scientific areas (e.g.…”

The path to VICTORy – a beginner's guide to success using commercial research antibodies

“…These include microcapillary digital western blots (WBs), high-density peptideand protein-display arrays, coupled immunoprecipitation/mass spectroscopy (IP/MS) and high multiplex IHC. However, a survey by the Global Biological Standards Institute (GBSI) suggested that only amongst well-established investigators did a majority even see a need to validate their cr-Abs (Freedman et al, 2016). Furthermore, not all cr-Ab suppliers have the best interests of researchers entirely at the focus of their operations: vials may lack active or indeed any reagent (Couchman, 2009) or may have forged labels (Cyranoski, 2017), ELISA kits may be non-selective , producer animal husbandry may be appallingly deficient (Lowe, 2016), and cr-Abs may bind molecules entirely divorced from the one advertised (Andersson et al, 2017;Gilda et al, 2015), to offer some examples.…”

“…The examples above emphasize the need to specifically design appropriate controls to confirm that any cr-Ab used is F4P in each and every experimental context it might be used in. For further details, the reader is referred to excellent discussions elsewhere (Freedman et al, 2016;Roncador et al, 2016;Torlakovic et al, 2015;Uhlén et al, 2016;Voskuil, 2014).…”

“…As discussed above, unequivocal cr-Ab identification, validation with appropriate controls and the full communication of these data are fundamental. Although many university biology courses are superb, it seems that only those researchers scared and scarred by experience routinely tend to be aware of this (Freedman et al, 2016). However, the issues we are facing in the use of cr-Abs are existential and we must address them (Baker, 2015b;Begley and Ellis, 2012;Goodman, 2018).…”

“…Hybridoma cell lines make single antibodies (Köhler and Milstein, 1975), and these lines can provide a reproducible and identifiable resource. They are moderately expensive to make, but they are in principle 'immortal' (Borrell, 2010;Freedman et al, 2016;Yu et al, 2015). However, rodent hybridomas frequently come from inbred murine populations with limited antibody diversity, which restricts cross-reactivity with mouse, a frequent object of analysis, and they can be destabilized by their unusual genetic complement (Barnes et al, 2003).…”