{"id":9026,"date":"2016-08-31T01:32:26","date_gmt":"2016-08-31T01:32:26","guid":{"rendered":"http:\/\/admin.acceleratingscience.com\/behindthebench\/?p=9026"},"modified":"2016-10-03T20:30:53","modified_gmt":"2016-10-03T20:30:53","slug":"the-path-to-antibody-specificity-confirmation","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/behindthebench\/the-path-to-antibody-specificity-confirmation\/","title":{"rendered":"The Path to Antibody Specificity Confirmation"},"content":{"rendered":"

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Editor’s note: research\u00a0antibodies are critical for innovation and discovery, but the use of incompletely characterized reagents can negatively impact the rigor and reproducibility of biomedical science, potentially leading to wasted time and resources. Comprehensive antibody validation standards and specificity confirmation will help researchers proceed with confidence, knowing they can count on the reagents they use.<\/em><\/p>\n

About Mathias Uhl\u00e9n: Dr. Uhl\u00e9n, Professor of Microbiology at Royal Institute of Technology, Sweden, is a noted protein biochemist, the founder of the Human Protein Atlas, and Chair of the International Working Group on Antibody Validation<\/a>.<\/em><\/p>\n

Mathias Uhl\u00e9n, Chair of the International Working Group on Antibody Validation<\/p><\/div>\n

Research antibodies are one of the workhorses that drive innovation and discovery across the biomedical sciences: on average, a typical scientist may perform more than three antibody-based assays each week. [BIOCOMPARE 2015] Frequent use of antibodies in the laboratory highlights the fundamental importance of antibody-based techniques for modern science, from the study of stem cells to investigation of cancer biology. Frequent antibody use also illustrates how the systematic application of incompletely characterized antibodies (and misuse of those that are well characterized) might quickly lead to confusion, misinterpretation of data, and irreproducibility in the scientific literature. These concerns (first ignited in 2012 by reports that only a small proportion of landmark pre-clinical studies in oncology could be reproduced by scientists at Amgen) have led to calls by the National Institutes for Health (NIH) and others to standardize the use of research reagents, including antibodies. [BAKER 2015B; BEGLEY 2012; COLLINS 2014]<\/p>\n

Recognition of potential pitfalls with antibody use is nothing new. As early as 2009, a study of 24 antibodies thought to specifically recognize different muscarinic receptor (MR) subtypes found that each antibody (with only 2 exceptions) recognized additional proteins expressed in tissues of corresponding MR-receptor-knockout mice. This finding led the authors to conclude that \u201creliable immunohistochemical localization of MR subtypes with antibodies is the exception rather than the rule.\u201d [JOSITCH 2009] \u00a0Published reports like this one (and likely, the personal experience of countless other investigators) have led to calls for universal standards to improve antibody specificity and reproducibility to ultimately enable more effective biomedical research. (Figure 2) <\/strong>[FASEB 2016; BORDEAUX 2010; BAKER 2015B]<\/p>\n

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Figure 2. Milestones in the Debate Surrounding Antibody Validation<\/p><\/div>\n

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Given the critical need for expanded antibody validation standards, including for antibody specificity confirmation, developing the underlying strategies used to confirm antibody specificity and improve reproducibility poses the next challenge. I would argue that the most successful approaches will adhere to four key attributes:<\/p>\n