The International Working Group on Antibody Validation (IWGAV), a completely independent group of international scientists with diverse research interests in the field of protein biology, today announced the publication of initial strategies made to address an important unmet need for antibody specificity, functionality and reproducibility in the online issue of Nature Methods. The IWGAV is the first initiative of the its size and scope to establish strategic strategies for antibody validation for antibody producers and users.
Although antibodies are the most frequently used tools in scientific research, there isn't a comprehensive scientific framework to the validation of antibody specificity across research applications. The use of poorly characterized antibodies has been estimated to cost $350 million annually in the United States alone and $800 million worldwide on account of failed or unreliable experiments.
"This publication is a first step towards the development of widely accepted standards for validating antibodies and ensuring good qualitywith the IWGAV. "
We look forward to receiving feedback in the broader community of antibody users, publishers, funding agencies and producers for helping strengthen this initial proposal and make sure the reliability of these essential tools of biomedical research."
The IWGAV identified five "conceptual pillars" to help antibody validation in specific research applications:
Genetic strategies: Study the relevant signal in control cells or tissues where the target gene has become knocked out or knocked down using techniques for example CRISPR/Cas or RNAi.
Orthogonal strategies: Use an antibody-independent method for quantification across multitudes of samples and after that examine the correlation relating to the antibody-based and antibody-independent quantifications.
Independent antibody strategies: Use several independent antibodies that recognize different epitopes on the target protein and confirm specificity via comparative and quantitative analyses.
Expression of tagged proteins: Customize the endogenous target gene to include sequences on an affinity tag or perhaps a fluorescent protein. The signal through the tagged protein may be correlated with detection through antibody-based methods.
Immunocapture followed by mass spectrometry (MS): Couple immunocapture, the process of isolating a protein from a solution through binding having a target-specific antibody, with MS analysis to spot proteins that interact directly using the purified antibody in addition to proteins that will form a fancy with the target protein.
While each of these conceptual pillars may provide proof of antibody specificity, the IWGAV recommends multiple pillars be utilized to claim a certain antibody has been well validated for use in a specific application. The manuscript in nature Methods also may include recommendations for producers and users to ensure antibody reproducibility over time.
The proposal, published in Nature Methods is intended to provide a scientific foundation for antibody validation and reproducibility standards that encourage the widespread adoption of comprehensive guidelines for antibody use. However, this may require continued input from your global research community and antibody producers. All these stakeholders will have important roles to learn in the adoption and implementation of the proposal.
More information: A proposal for validation of antibodies, Nature Methods, nature.com/articles/doi:10.1038/nmeth.3995
Souce: NovoPro 2016-09-09