A variety of issues-from production variations and storage conditions to improper validation techniques-can jeopardize even the best-designed biomedical experiments involving an antibody. Research antibodies must demonstrate specificity, selectivity and reproducibility in the application or assay for which they are used. Unfortunately, there is no simple solution to ensure this happens in every experiment across all labs.
To determine a greater understanding of current research antibody usage and validation practices, the Global Biological Standards Institute (GBSI) designed and fielded an online, anonymous survey in 2015. Key findings from the survey published in BioTechniques, indicate that many scientists-particularly junior researchers-are not receiving the training they need in the proper use and validation of research antibodies.3 The survey highlights a potentially dangerous perception among many researchers that antibody validation delays, rather than facilitates, research progress.3
Responses from more than 500 scientists within the life science research community who responded to the survey showed that, although 70% of respondents validate or have commercial antibodies validated, just 43% of researchers with fewer than five years’ experience reported validating purchased antibodies-and only 22% reported validating antibodies produced in-house. Still more troubling, the survey found that a third of these future lab heads do not validate at all.
Significantly, less than half (44%) of the respondents reported receiving specific training on validation and how to validate research antibodies for specific applications. When assessing barriers to validating antibodies in their research, a clear trend has emerged based on years of experience-more than 25% of researchers with fewer than five years of antibody use indicated that they “do not see the necessity” of performing antibody validation. Other perceived barriers included laboratorians feeling that it “takes too much time” (71%) and “delays my research” (51%).
The research antibody survey clearly identified a need to improve the skills and proficiency of junior scientists in application-specific antibody validation and to develop validation standards that are widely accepted. The biomedical research community must commit sufficient time, resources and expertise to educating and training scientists in best practices for antibody-based experiments and reporting, as well as application-specific validation.3The GBSI survey also indicated that journals should require authors to identify and describe all antibodies used in a study as an explicit condition of publication.4 Essential information included the name of the vendor or individual that supplied the antibody, the immunogen used to generate the antibody, the nature of the antibody preparation (e.g., polyclonal or monoclonal), catalog and lot number and validation status-including primary validation data if not previously published.
Because vendor catalog numbers are not stable over time as products are discontinued or sold,5 GBSI also recommended linking each antibody to one or more online registries or databases and a dedicated bioresource portal like the eagle-i Network6 or the Resource Identification Portal.7 In addition, recent NIH initiatives to improve the rigor and reproducibility of funded research clarified and revised grant application instructions and reviewed criteria as of January 25, 2016. They included expectations, but do not explicitly require that key biological resources such as antibodies will be “regularly authenticated to ensure their identity and validity for use in the proposed studies.”
As stated in the GBSI survey summary, “Despite their prolific use in the laboratory, however, there are no standardized and widely accepted guidelines for how to validate antibodies prior to use in preclinical research, although some approaches have been proposed.”3
In order to accelerate the successful translation of bench-side research breakthroughs into approved diagnostics and therapies, standards and best practices must be established around the development, commercial availability and widespread use of assay-specific validated antibodies in biomedical research. It is important that the life science research community create validation tools, standards and methods that are viewed not as obstacles, but instead aids to improving research reproducibility.
Additionally, GBSI is hosting a 3-day workshop with the express purpose of developing research antibody standards. The Antibody Validation: Standards, Policies, and Practices workshop takes place from September 25th-27th, 2016.
Follow the discussion at #AbValidate and @gbsiorg.
- Begley, C.G., and J.P. Ioannidis. 2015. Reproducibility in science: improving the standard for basic and preclinical research. Circ. Res 116:116-126.
- Freedman, L.P., I.M. Cockburn, and T.S. Simcoe. 2015. The economics of reproducibility in preclinical research. PLoS Biol 13:e1002165.
- Freedman, L. P. et al. BioTechniques 61. (2016).
- Schonbrunn, A. 2014. Editorial: antibody can get it right: confronting problems of antibody specificity and irreproducibility. Mol Endocrinol 28:1403-1407.
- Voskuil, J. 2014. Commercial antibodies and their validation. F1000Res 3:232.
- Vasilevsky, N.A., M.H. Brush, H. Paddock, L. Ponting, S.J. Tripathy, G.M. LaRocca, and M.A. Haendel. 2013. On the reproducibility of science: unique identification of research resources in the biomedical literature. PeerJ 1:e148.
- Bandrowski, A., M. Brush, J.S. Grethe, M.A. Haendel, D.N. Kennedy, S. Hill, P.R. Hof, M.E. Martone, et al. 2016. The Resource Identification Initiative: a cultural shift in publishing. J Comp Neurol 524:8-22.