The European antibody network's practical guide to finding and validating suitable antibodies for research

Abstract

Antibodies are widely exploited as research/diagnostic tools and therapeutics. Despite providing exciting research opportunities, the multitude of available antibodies also offers a bewildering array of choice. Importantly, not all companies comply with the highest standards, and thus many reagents fail basic validation tests. The responsibility for antibodies being fit for purpose rests, surprisingly, with their user. This paper condenses the extensive experience of the European Monoclonal Antibody Network to help researchers identify antibodies specific for their target antigen. A stepwise strategy is provided for prioritising antibodies and making informed decisions regarding further essential validation requirements. Web-based antibody validation guides provide practical approaches for testing antibody activity and specificity. We aim to enable researchers with little or no prior experience of antibody characterization to understand how to determine the suitability of their antibody for its intended purpose, enabling both time and cost effective generation of high quality antibody-based data fit for publication.

Challenges in Developing a Biochip for Intact Histamine Using Commercial Antibodies

“…Several authors have addressed the issue and claimed for better characterization and profound information on the specificity and cross-reactivity of the antibodies [19][20][21][22][23][24][25][26][27][28]. The antibodies in the market are often insufficiently characterized and furthermore they might possess batch-to-batch variation, usually in much larger extent with polyclonal antibodies compared to monoclonal.…”

“…At present, some databases such as antibodypedia.com or pabmabs.com had no entries for histamine specific antibodies. The common databases should prevent the overlapping supply between the different vendors; nonetheless, the same antibody can be sold by different providers just with different labels and product number and users might accidentally buy the same reagent from different distributors [19,28]. Thus, each antibody should be adequately identified and linked to a code that should be kept unaltered by users and vendors.…”

“…The recommended applications are mainly immunohistochemistry (IHC) or immunocytochemistry (ICC), but also western blot, EIA and ELISA. Recently, several studies have pointed out the quality issues and traceability problem concerning commercial antibodies and the lack of quality in the antibody market has been widely recognized by the scientific community [19][20][21][22][23][24][25][26][27][28]. Often, the characterization and specifications provided by the manufacturers are somewhat incomplete and, furthermore, the authors often fail to identify the exact antibody used [28][29][30] for other researchers to be able to purchase the same product.…”

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

“…There are currently no standardized and widely accepted guidelines for antibody validation, although some approaches have been proposed, such as by Bordeaux et al (13) and more recently by Roncador et al (14). In 2014, the Global Biological Standards Institute (GBSI) formed a Research Antibodies and Standards Task Force to examine: ( i ) specific areas where standards for research antibody characterization and validation are urgently needed; ( ii ) what types of standards could be designed and implemented to address these needs; and ( iii ) how these recommendations can be implemented to advance the availability and use of reliable, high quality antibodies across the biomedical research community.…”

Determination of equilibrium dissociation constants for recombinant antibodies by high-throughput affinity electrophoresis

“…Immunoreagents are notorious for variation in quality and performance12. Differences in specificity, binding affinity, and even lot-to-lot performance are widely reported, negatively impacting resources and reporting3.…”